Next Article in Journal
Improving HCV Screening in Addiction Care Centers with Plasma Separation Cards
Previous Article in Journal
Acute Influenza Virus-Associated Encephalitis and Other Neurological Complications in Severe Hospitalized Laboratory-Confirmed Influenza Cases—Catalonia 2010–2020
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Pathogens
Volume 14
Issue 3
10.3390/pathogens14030238
pathogens-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Rapid Detection of Panton–Valentine Leukocidin Production in Clinical Isolates of Staphylococcus aureus from Saxony and Brandenburg and Their Molecular Characterisation †

by
Elke Müller
1,2,‡,
Stefan Monecke
1,2,*,‡,
Marc Armengol Porta
3,
Marco Vinicio Narvaez Encalada
4,
Annett Reissig
1,2,
Lukas Rüttiger
5,
Percy Schröttner
6,7,
Ilona Schwede
8,
Hans-Herman Söffing
5,
Alexander Thürmer
9 and
Ralf Ehricht
1,2,10
1
Leibniz Institute of Photonic Technology (Leibniz-IPHT), Leibniz Center for Photonics in Infection Research (LPI), Germany and InfectoGnostics Research Campus, 07745 Jena, Germany
2
InfectoGnostics Research Campus, Centre for Applied Research, 07745 Jena, Germany
3
Medizinisches Labor Ostsachsen MVZ GbR, 01067 Dresden, Germany
4
Städtisches Krankenhaus Dresden, 01067 Dresden, Germany
5
Senova Gesellschaft für Biowissenschaft und Technik mbH, 99427 Weimar, Germany
6
Institute for Medical Microbiology and Virology, Faculty of Medicine and University Hospital “Carl Gustav Carus”, Technische Universität Dresden, 01307 Dresden, Germany
7
Institute for Clinical Chemistry and Laboratory Medicine, Faculty of Medicine and University Hospital “Carl Gustav Carus”, Technische Universität Dresden, 01307 Dresden, Germany
8
IMD Labor Oderland GmbH, 15230 Frankfurt (Oder), Germany
9
MVZ Medizinische Labore Dessau Kassel GmbH, 06847 Dessau, Germany
10
Institute of Physical Chemistry, Friedrich Schiller University Jena, 07745 Jena, Germany
*
Author to whom correspondence should be addressed.
†
A preliminary analysis of the dataset was presented as poster at the annual conference of the German Society for Microbiology and Hygiene in Würzburg, Würzburg, Germany, 2–5 June 2024.
‡
These authors contributed equally to this work.
Pathogens 2025, 14(3), 238; https://doi.org/10.3390/pathogens14030238
Submission received: 27 January 2025 / Revised: 24 February 2025 / Accepted: 27 February 2025 / Published: 1 March 2025
(This article belongs to the Section Bacterial Pathogens)
Browse Figure
Versions Notes

Abstract

:
Panton–Valentine leukocidin (PVL) is a staphylococcal toxin associated with chronic/recurrent skin and soft tissue infections (SSTIs) and necrotizing pneumonia. Its detection in clinical isolates of Staphylococcus aureus warrants aggressive therapy and infection control measures. However, PVL detection relies on molecular methods of limited use, especially in outpatient or resource-poor settings. In order to aid the development of a lateral flow (LF) test for PVL, clinical isolates from SSTIs were collected in 2020/21 at three laboratories in two cities in the Eastern part of Germany. After the exclusion of duplicate and serial isolates, 83 isolates were eligible. These were tested using an experimental LF test for PVL production. They were also characterized using DNA microarrays, facilitating the detection of virulence and resistance markers as well as the assignment to clonal complexes and epidemic/pandemic strains. Thirty-nine isolates (47%) were PVL-positive, and the LF results were in 81 cases (97.6%) concordant with genotyping. One false-positive and one false-negative case were observed. This translated into a diagnostic sensitivity of 0.974 and a diagnostic specificity of 0.977. The most common PVL-positive MSSA lineages were CC152 (n = 6), CC121 (n = 4), and CC5 and CC30 (each n = 2). Thirty isolates (36%) were mecA-positive. The MRSA rate among PVL-negatives was 20% (nine isolates), but among the PVL-positives, it was as high as 54% (n = 21). The most common PVL-MRSA strains were CC398-MRSA-VT (n = 5), CC5-MRSA-IV “Sri Lanka Clone” (n = 4), CC8-MRSA-[mec IV+Hg] “Latin American USA300” (n = 4), and CC22-MRSA-IV (PVL+/tst+) (n = 2). While the PVL rate was similar just like the German isolates from a previous study a decade before, the MRSA rate among PVL-positives was clearly higher. All PVL-MRSA strains detected, as well as the most common methicillin-susceptible lineage (CC152), are known to be common locally in other parts of the world, and might, thus, be regarded as travel-associated. Therefore, patients with suspected PVL-associated disease should be asked for their history of travel or migration, and, in case of hospitalization, they should be treated as MRSA cases until proven otherwise.

1. Introduction

Staphylococcus (S.) aureus is a ubiquitous Gram-positive bacterium that colonizes or infects humans as well as a wide variety of mammal and bird species. It can cause various skin and soft tissue infections (SSTIs), osteomyelitis, pneumonia, sepsis, and toxin-mediated conditions, such as toxic shock syndrome and food intoxications. Many strains of this bacterium carry resistance genes on mobile genetic elements, with the most notable example being mecA (and/or the related mecC, which, however, has never been found in PVL-positive strains), localized on SCCmec elements [1,2,3,4]. This gene encodes an alternative β-lactam-binding protein and, thus, causes resistance to most β-lactam antibiotics. Strains that carry it are named methicillin-resistant S. aureus, or MRSA, as methicillin is an indicator substance for this phenomenon. S. aureus strains also can variably harbour mobile genetic elements that include genes for various toxins and virulence factors.
One virulence factor that often plays a role in the development of SSTIs and pneumonia in humans is the Panton–Valentine leukocidin (PVL, [5,6,7,8]). It is encoded by two neighbouring genes located together on prophages [9,10,11] integrated invariably at the same position into the bacterial chromosome (lukS-PV and lukF-PV, GenBank BA000033.2:MW1378 and MW1379; [12]). This genetic arrangement allows for the horizontal transfer of the PVL toxin between different S. aureus strains through a process called lysogenic conversion. When the prophage is induced, it can replicate lyse, the host cell, and infect other S. aureus bacteria, potentially transferring the PVL genes to new staphylococcal cells. This mechanism of transmission contributes to the spread of PVL across diverse and rather unrelated lineages of S. aureus, and even into related staphylococcal species [13]. Together, the molecules of the two components of leukocidin genes form polymeric pores in leukocyte membranes. This kills these cells in a highly specific manner resulting in the release of cytokines [8,14] which, in turn, attracts more leukocytes. While PVL is rather specific for human leukocytes [15], they are structurally and functionally similar to leukocidins, and can be detected in S. aureus strains from various animal species. LukM/lukF-P83 [8] is known for ruminant strains causing mastitis and for a strain associated with dermatitis in tree squirrels [16,17]. Another set of leukocidin genes, called lukP/lukQ, has been identified in S. aureus strains isolated from horses [18,19] while small ruminants harbour strains with a chimeric leukocidin derived from lukP/lukQ and lukM/lukF-P83 [19]. Yet, another leukocidin (lukF/S-BV) which is closely related to PVL was recently discovered in S. aureus from Eurasian beavers [20]. As all are carried on prophages, this suggests that phage infections of S. aureus may predetermine both host specificity and clinical manifestations of S. aureus infection in its vertebrate host.
PVL-associated diseases in humans occur particularly in immunocompetent young, previously healthy patients. Clinically, they manifest as recurrent or chronic SSTIs, abscesses, furunculosis, as well as so-called “spider-bite lesions”. Case clusters or outbreaks frequently affect families, people who are confined to close proximity to each other, such as inmates of prisons or recruits in army barracks, or those who share possibly contaminated items such as towels, diving masks, breathing devices, etc. PVL-associated disease also includes abscessing or necrotizing pneumonia that can be a complication of viral respiratory tract infections and has a mortality rate of as high as 40% [5,7,21].
While PVL is present only in about a small percentage of S. aureus isolates [22], it can be detected much more frequently in isolates originating from SSTIs [5,23]. There used to be large differences in the prevalence of PVL depending on geographical regions, with PVL-positive strains being common in, for instance, Northern America, India, the greater Middle East, sub-Saharan Africa [24], and Australia while being rare in Northern and Central Europe [22], although this might change with the spread of PVL-MRSA. Most interestingly, the prevalence of anti-PVL antibodies exhibits a notable north–south gradient, with significantly higher levels observed in African populations compared to European ones [25]. This serological difference reflects the varying exposure to PVL-positive S. aureus strains across geographical regions, and it might also contribute to the apparent rarity of severe/fatal PVL infections in regions where PVL-positive staphylococci are common.
In the 1940s and 1960s, and from the mid-1990s to the present day, there have been worldwide clusters of furunculosis caused by PVL-positive strains. While the first known strains were sensitive to all antibiotics (typical representatives are the laboratory strains ATCC23925 [26] and “Oxford Staphylococcus” [27]), the strains that dominated in the 1960s (phage type 80/81) and thereafter were mostly penicillinase-positive. Epidemiological studies from many countries around the world show that PVL-positive MRSAs are currently spreading. There are distinct strains that predominate in different geographic regions. In particular, the strain known as “USA300” became very common in the USA during the last decades, with approximately 60% of all community-acquired skin and soft tissue infections caused by S. aureus being attributable to this strain [28,29]. Other strains originate from the Indian subcontinent [30,31,32,33] or Australia [34,35,36,37]. In Europe, a wide variety of different strains of PVL-MRSA occur, and many cases have travel histories or family ties in regions where PVL-MRSA is more common [38,39,40,41,42].
Because of the higher virulence of PVL-positive strains, a more aggressive approach to therapy is warranted. This includes a more liberal administration of antibiotics in addition to incision and drainage of abscesses, and it also includes infection control and eradication measures akin to those applicable in the case of MRSA colonization/infection [43,44]. PVL-MRSA, obviously, are more complicated to treat, and for that reason, it is discussed whether infections with PVL-MRSA should be declared a notifiable disease. In the German state of Saxony, this is already the case [45,46]. However, infection control measures are hindered by the lack of a convenient diagnostic tests. PVL detection is currently essentially limited to molecular methods. These normally are performed only in specialized laboratories with dedicated hardware and trained personnel, and they require complex sample preparation. Most cases, however, are seen in the outpatient setting, where molecular tests are not readily available. A referral of isolates to reference laboratories usually requires too much time until results are available, so that these results do not have a significant impact on the case management anymore. In addition, costs play a major role as general practitioners have limited budgets for laboratory tests while many patients come from marginalized sections of the population, such as prison inmates, substance users or refugees.
For these reasons, efforts are underway to develop a lateral flow assay for the detection of PVL from S. aureus cultures. Lateral flow tests have achieved some popularity during the COVID-19 pandemic, so a short description of the principle should be sufficient. The test device contains target-specific antibodies, IgG antibodies coated to particles, and other target-specific IgG antibodies coated on a membrane. During testing, the molecules from the sample react with the antibodies coating the particles. The mixture migrates upward on the membrane chromatographically, by capillary force, to react with captured antibodies on the membrane and generate a coloured line. Its presence in the test region indicates a positive result, while its absence indicates a negative result. To serve as a procedural control, a coloured line will always appear in the control line region.
In order to aid the development of a lateral flow test for PVL, clinical isolates from SSTI were collected in 2020/21 at two laboratories in Eastern parts of Germany. Isolates were tested using an early experimental stage of the LF assay, and they were genotyped in parallel using a DNA-based microarray assay that allowed not only to confirm or to rule out the presence of PVL genes and mecA, but also to assign a given isolate to clonal complexes (CC, i.e., phylogenetic lineages within S. aureus) and known epidemic strains.

2. Materials and Methods

2.1. Isolates

Isolates were collected in 2020/21 at two laboratories in Dresden, Saxony, Germany, and at one in Frankfurt/Oder, Brandenburg, Germany. Clinical partners were advised to submit isolates originating from furunculosis or carbuncles, cutaneous abscesses, other conspicuous or severe skin and soft tissue infections, such as mastitis or necrotizing fasciitis, chronically purulent and painful “spider bites”, recurrent or chronic skin- and soft tissue infections (with the exception of diabetic foot ulcers), as well as isolates from cases of necrotizing community-acquired pneumonia, including cases associated with influenza. After the exclusion of duplicate and serial isolates, eighty-three isolates were eligible for study. Twenty-six originated from a communal/city hospital in Dresden (Dresden Friedrichstadt), thirty from the University Hospital of Dresden, and twenty-seven from a diagnostic laboratory serving healthcare providers in and around Frankfurt/Oder.

2.2. Antibodies and Lateral Flow Test Productions

Antibodies were generated using the mouse cell hybridoma method, according to Kohler and Milstein [47] using the recombinant lukF-PV gene product as an antigen. The manufacturing of the lateral flow tests was performed in Senova, Weimar, Germany.

2.3. Lateral Flow Test

S. aureus isolates were incubated on a Columbia blood agar overnight for up to 24 h at 37 °C ± 2 °C. If contaminations or mixed cultures were suspected, cultures were re-cloned. Catalase and coagulase/clumping factor tests or species identification by MALDI-TOF were performed to confirm S. aureus. One inoculation loop (ca. 1 µL) full of culture material was inoculated into 300 µL of a buffer (phosphate-buffered saline, PBS, plus 0.05% v/v Tween20) and the mixture was shortly vortexed. Then, it was centrifuged in order to sediment the cells (30 s at 2000× g). In total, 100 µL of the supernatant was pipetted onto the sample well of the test device, which was then incubated for 10 min at room temperature. The results were visually read: the presence of both test and control lines was considered positive, while the presence of the control line alone was considered negative. An absence of the control line was interpreted as invalid. The test results for the individual isolates are shown in Supplemental file S1.

2.4. Microarray Procedures

The molecular characterization was carried out using the InterArray Genotyping Kit S. aureus (InterArray GmbH, Bad Langensalza, Germany) for the detection of species markers as well as of virulence and resistance genes. Probes, primers, the test principle, and procedures were previously described in detail ([48,49,50]; https://doi.org/10.1371/journal.pone.0017936.s001; accessed on 20 February 2025). In short, a multiplexed primer elongation reaction was performed on sample DNA from pure culture amplifying and labelling specific fragments of the target genes. These were hybridized to the microarray carrying specific probes in a predefined pattern. Hybridization was detected using an enzymatic precipitation reaction triggered by the labels incorporated into the amplicons. The microarray harboured probes for both, lukS-PV and lukF-PV, thus facilitating the molecular confirmation of the lateral flow immunoassay as well as the detection of other virulence genes, resistance genes, and markers that were used for the assignment to CCs and strains. Microarray images were taken, and analysis was carried out using a dedicated reader and software (InterArray GmbH; software versions 1.0.7 gamma to 1.1.8). The full array data are shown in Supplemental file S1.

3. Results

One hundred isolates were collected, but after the exclusion of duplicate and serial isolates, eighty-three isolates were finally eligible for the study. Thirty-nine isolates (47%) were PVL-positive. Microarray-based genotyping revealed one false-negative result and one (weakly) false-positive result. This translated into a diagnostic sensitivity of 0.974, a diagnostic specificity of 0.977, a positive prediction value of 0.974, a negative prediction value of 0.977, and an accuracy of 0.976.
The PVL- and mecA-status, as well as CC and strain affiliations of the isolates, are summarized in Table 1. Thirty isolates (36.1%) were mecA-positive. In total, 35 isolates (42.2% of the total) were PVL-negative MSSA, and 9 isolates (10.8%) were PVL-negative MRSA. The MRSA rate among PVL-negatives was thus 20.5%. The most common lineages among PVL-negative MSSA were CC15, CC22, CC30, and CC45. PVL-negative MRSA included a variety of different strains; see Discussion. Eighteen isolates (21.7% of the total) were PVL-positive MSSA and twenty-one (25.3%) were PVL-positive MRSA. The MRSA rate among PVL-positives was as high as 54% (n = 21).
The most common PVL-positive MSSA lineages were CC152 (n = 6), CC121 (n = 4), and CC5 and CC30 (each n = 2). The most common PVL-MRSA strains were CC398-MRSA-VT (n = 5), CC5-MRSA-IV “Sri Lanka Clone” (n = 4), CC8-MRSA-[IV+Hg] “Latin American USA300” (n = 4), and CC22-MRSA-IV (PVL+/tst+) (n = 2). Other PVL-MRSA were CC1-MRSA-[V+fus+ccrAB1], CC8-MRSA-[IV+ACME] “USA300”, CC30-MRSA-IV “WSPP Clone”, CC88-MRSA-IV, CC152-MRSA-IV, and –[V+fus], with one isolate each (see Discussion for details).
The alternate gene for methicillin resistance, mecC, was not found. The SCCmec-borne gene for fusidic acid resistance, fusC, was detected in nine isolates, all of them being MRSA, two being PVL-positive. The mupirocin resistance gene mupA was found twice, in a PVL-MRSA (of the “USA300” strain) and in a PVL-negative MSSA. The prevalence rates for genes responsible for resistance to antibiotic compounds as detected by microarray are listed in Table 2. Given that first-line antibiotics for the treatment of PVL-associated SSTI include oral trimethoprim-sulfamethoxazole, clindamycin, and doxycycline [44], the high rates of carriage of relevant resistance genes, especially among PVL-MRSA (dfrA, 19%; tet-genes, 47.6%; erm-genes, 52.4%), warrant attention.

4. Discussion

The experimental performance, as confirmed by the genotypic experiments of the lateral flow assay, was fairly sufficient, given that this was an experimental, pre-production batch. A single false-negative result might be attributed to poor or absent in vitro toxin production. It is known that in vitro expression levels of the toxin vary widely [51,52], and that, in addition to culture conditions [53], mutations in regulatory genes might affect it [52,54]. Thus, it is not surprising that an isolate might be found in which lukS/F-PVL genes were present, although the protein was undetectable. The false-positive result remained unexplained. Further studies might focus on culture conditions, i.e., on growth media composition and supplements, as well as on optimization of the assay and its procedures. In general, the lateral flow test provides a suitable tool for a rapid and inexpensive detection of PVL by clinical isolates of S. aureus. This might appeal to laboratories that do not routinely perform molecular tests. It also might shorten the time until a test result is available to the care provider, especially in settings where it was otherwise necessary to ship suspected isolates to central reference laboratories.
A very similar study was conducted by the authors roughly a decade ago [55], and a direct comparison of the present results to the ones of the German branch of this previous study is interesting (Figure 1).
Then, fifty-six isolates were characterized that were collected in Dresden or submitted by cooperating centres to the Dresden laboratory, using the same inclusion criteria. Twenty-six (46.4%) of these isolates were PVL-negative MSSA, and four (7.1%) were PVL-negative MRSA. Twenty isolates (35.7%) were PVL-positive MSSA, and six (10.7%) proved to be PVL-positive MRSA. Thus, it can be assumed that the “ecological niche” for PVL-positives remained unchanged during that decade. However, a clear shift occurred from PVL-positive MSSA to PVL-positive MRSA, whose share of the total increased from about 10% to about 25%. This is an interesting observation. From the eastern part of Germany, no large outbreak or epidemic situation of community-associated MRSA (ca-MRSA) and/or PVL-MRSA has been described to date. The only major outbreak in Germany of such a clone was observed in the early 2000s in Bavaria [56]. Since then, PVL-positive MRSA has been uncommon, and there is no single predominant clone such as “USA300” in the USA [57,58,59,60]. Contrarily, there was, for decades, a number of different, sporadic strains, which can easily be explained by random importations affecting travellers and possibly immediate contacts, such as family members, in absence of an ongoing epidemic transmission in the local community [57,58,59,60].
The most common methicillin-susceptible/PVL-positive lineages in this study were CC121 and CC152. The former can be found essentially anywhere in the world. The latter is known to be common in Sub-Saharan Africa [61,62,63,64,65,66].
All PVL-MRSA strains detected are known to locally predominate in other parts of the world, and thus, all might be regarded as travel-associated. The most common PVL-MRSA in this study was CC398-MRSA-VT (PVL+). While PVL-negative CC398-MRSA became well-known as livestock-associated MRSA lineage in Western Europe, PVL-positive specimens of CC398 were frequently described from Southeast Asia and from travellers with a corresponding travel history. Touristic travel might, here again, have played a role in the importation of that strain, as well as the fact that the Eastern German Federal States are home to a large community of Vietnamese descent.
Some of the strains observed indicate epidemiological connections to the Middle East and could be regarded as associated either with travel to popular tourist destinations or to migration from Middle Eastern zones of political upheaval. These strains include CC1-MRSA-[V+fus+ccrAB1 (PVL+), which has been observed by the authors as well as by others in the greater Middle East, especially Egypt [67,68]. CC22-MRSA-IV (PVL+/tst+) was apparently first observed in Iran [69], but more recently, it can be found elsewhere in the Gulf region [70], and it is spreading into Asia where it has already been detected in China [71] and Nepal [72]. CC88-MRSA-IV (PVL+) and CC152-MRSA-[V/VT+fus] (PVL+) also have been observed in the Gulf region and Egypt [67,68]. C152-MRSA-IV is a sporadic strain with anecdotal evidence pointing towards Eastern Africa (GenBank accession numbers CP141387.1, CP141398.1, CP141523.1) and the Middle East [70].
CC8-MRSA-[IV+Hg] (PVL+) “Latin American USA300” occurs in Southern America and Spain [73,74]. CC8-MRSA-[IV+ACME] (PVL+) “USA300” is common in Northern America and, to a lesser extent, in other Anglophone countries of the world. CC30-MRSA-IV (PVL+) “WSPP Clone” originated from the West Pacific region [75,76] but was meanwhile observed, although sporadically, in many countries. CC5-MRSA-IV (PVL+) “Sri Lanka Clone” is an emerging clone that was first described from Sri Lanka but that recently caused outbreaks across several European countries [77,78]. For all these strains, travel and tourism might have played a role in the importation of these strains into the study settings.
Regarding PVL-negative MRSA strains, one isolate belonged to CC22-MRSA-IV. This strain (“UK EMRSA15” or locally known as “Barnim epidemic strain”, [79]) is a common healthcare-associated MRSA strain in Eastern Germany [58]. One isolate was the “European CC1-MRSA-IV strain”, a strain originating from Southeastern Europe that spread to several Central/Western European countries, possibly aided by its ability to evade a frequently used molecular screening test [80,81,82]. The detection of as many as three isolates of CC5-MRSA-[IV+fus+ccrAB], “Maltese Clone”, all from Frankfurt/Oder, is somewhat startling. This strain was first described in Malta [83], which is a tourist destination, but it was also observed in the Kingdom of Saudi Arabia [84,85] and the UAE [86], possibly indicating a wider distribution in the Middle East. Thus, for this strain, as well as for the other fusC-positives (because fusC in MRSA appears to be common in the greater Middle East [67,68,87,88,89,90,91]), a connection to touristic travel and/or migration might be assumed.
Therefore, patients with suspected PVL-associated disease, and patients with any “ca-MRSA”—regardless of PVL-status—should be asked for their travel histories. Furthermore, patients with suspected PVL-associated disease should, in case of hospitalization, be treated as MRSA cases until proven otherwise, because the odds of indeed carrying MRSA are above 50%.
Given the increase in PVL-MRSA and their clinical relevance, a mandatory notification to public health authorities should be considered. In Saxony, PVL-MRSA infections are already a notifiable disease [45,46], but this is not the case in other German Federal States. The main obstacle to stringent surveillance of PVL-positive S. aureus and PVL-MRSA is currently the lack of a quick and cheap assay for PVL. After all, most cases are seen in outpatient settings rather than in tertiary care centres, so that a lack of availability of laboratory infrastructure and molecular diagnostics impedes testing for PVL genes. An easy-to-use non-molecular test such as a lateral flow assay out of a primary culture might thus prove very helpful.

5. Conclusions

In conclusion, a lateral flow test for a time- and cost-efficient detection of PVL in overnight cultures of S. aureus might prove helpful to clinical laboratories, especially in settings where molecular tests are not feasible and where shipment of isolates to central reference laboratories might cause undue delays. This might aid in making timely decisions on case management and infection control measures. Patients with suspected PVL-associated disease should be asked for a history of travel or migration because all PVL-MRSA strains observed, as well as the most common MSSA lineage, are known to be locally common in other parts of the world. More than half of PVL-positive isolates were MRSA. In case of hospitalization, patients with suspected PVL-associated disease should, therefore, be treated as MRSA cases until proven otherwise.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/pathogens14030238/s1, Supplemental File S1: Lateral flow and microarray test results.

Author Contributions

Conceptualization, S.M., H.-H.S. and R.E.; methodology, E.M., S.M., L.R., H.-H.S. and R.E.; investigation, E.M., M.A.P., M.V.N.E., A.R., L.R., P.S., I.S. and A.T.; resources, M.A.P., M.V.N.E., P.S., I.S., A.T., L.R. and H.-H.S.; data curation, E.M. and S.M.; writing—original draft preparation, S.M.; writing—review and editing, S.M., E.M. and R.E.; visualization, S.M.; supervision, R.E.; project administration, S.M., R.E. and H.-H.S.; funding acquisition, R.E. and H.-H.S. All authors have read and agreed to the published version of the manuscript.

Funding

We acknowledge the German Federal Ministry for Economic Affairs and Energy for supporting the development of the PVL antibodies and of the rapid assay within the framework of the INNO-KOM project (49MF180153).

Institutional Review Board Statement

Not applicable. Isolates originated from routine laboratory diagnostics and no samples were purposefully taken for the study.

Informed Consent Statement

Not applicable. No patient data were stored, processed or shared among the authors for study purposes.

Data Availability Statement

All relevant data are provided as Supplementary Materials. A preliminary analysis of the dataset was presented as poster at the annual conference of the German Society for Microbiology and Hygiene in Würzburg, 2024 [92].

Acknowledgments

We acknowledge the support of Peter Miethe (Posanova GmbH) and Anne-Marie Gad, Friedrich Scholz (Senova GmbH, Vellap GmbH) for the supply of the PVL antibodies and the lateral flow immunoassay. We thank the team at FZMB GmbH Erfurt and Biotech Rabbit GmbH Berlin for the generation and production of the antibodies and the teams in the clinical laboratories for collecting and providing isolates.

Conflicts of Interest

Lukas Rüttiger and Hans-Herman Söffing are employees of Senova GmbH. Ilona Schwede is an employee of IMD Labor Oderland GmbH. Alexander Thürmer is an employee of MVZ Medizinische Labore Dessau Kassel GmbH. The other authors declare that the study was conducted in the absence of any commercial or financial relationships that might pose a conflict of interest.

Abbreviations

The following abbreviations are used in this manuscript:
CCClonal complex
MALDI-TOFMatrix assisted laser desorption ionization (time-of-flight)
MRSAMethicillin-resistant Staphylococcus aureus
MSSAMethicillin-susceptible Staphylococcus aureus
PVLPanton–Valentine leukocidin
SSTISkin and soft tissue infection(s)
STSequence type (as defined by Multilocus Sequence Typing)

References

  1. Ito, T.; Katayama, Y.; Asada, K.; Mori, N.; Tsutsumimoto, K.; Tiensasitorn, C.; Hiramatsu, K. Structural comparison of three types of staphylococcal cassette chromosome mec integrated in the chromosome in methicillin-resistant Staphylococcus aureus. Antimicrob. Agents Chemother. 2001, 45, 1323–1336. [Google Scholar] [CrossRef] [PubMed]
  2. Ito, T.; Ma, X.X.; Takeuchi, F.; Okuma, K.; Yuzawa, H.; Hiramatsu, K. Novel type V staphylococcal cassette chromosome mec driven by a novel cassette chromosome recombinase, ccrC. Antimicrob. Agents Chemother. 2004, 48, 2637–2651. [Google Scholar] [CrossRef]
  3. Ito, T.; Okuma, K.; Ma, X.X.; Yuzawa, H.; Hiramatsu, K. Insights on antibiotic resistance of Staphylococcus aureus from its whole genome: Genomic island SCC. Drug Resist. Updat. 2003, 6, 41–52. [Google Scholar] [CrossRef]
  4. IWG-SCC. Classification of staphylococcal cassette chromosome mec (SCCmec): Guidelines for reporting novel SCCmec elements. Antimicrob. Agents Chemother. 2009, 53, 4961–4967. [Google Scholar] [CrossRef] [PubMed]
  5. Lina, G.; Piemont, Y.; Godail-Gamot, F.; Bes, M.; Peter, M.O.; Gauduchon, V.; Vandenesch, F.; Etienne, J. Involvement of Panton-Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clin. Infect. Dis. 1999, 29, 1128–1132. [Google Scholar] [CrossRef]
  6. Panton, P.; Valentine, F. Staphylococcal toxin. Lancet 1932, 1, 506–508. [Google Scholar] [CrossRef]
  7. Labandeira-Rey, M.; Couzon, F.; Boisset, S.; Brown, E.L.; Bes, M.; Benito, Y.; Barbu, E.M.; Vazquez, V.; Hook, M.; Etienne, J.; et al. Staphylococcus aureus Panton-Valentine leukocidin causes necrotizing pneumonia. Science 2007, 315, 1130–1133. [Google Scholar] [CrossRef] [PubMed]
  8. Kaneko, J.; Kamio, Y. Bacterial two-component and hetero-heptameric pore-forming cytolytic toxins: Structures, pore-forming mechanism, and organization of the genes. Biosci. Biotechnol. Biochem. 2004, 68, 981–1003. [Google Scholar] [CrossRef]
  9. Kaneko, J.; Kimura, T.; Kawakami, Y.; Tomita, T.; Kamio, Y. Panton-valentine leukocidin genes in a phage-like particle isolated from mitomycin C-treated Staphylococcus aureus V8 (ATCC 49775). Biosci. Biotechnol. Biochem. 1997, 61, 1960–1962. [Google Scholar] [CrossRef]
  10. Kaneko, J.; Kimura, T.; Narita, S.; Tomita, T.; Kamio, Y. Complete nucleotide sequence and molecular characterization of the temperate staphylococcal bacteriophage phiPVL carrying Panton-Valentine leukocidin genes. Gene 1998, 215, 57–67. [Google Scholar] [CrossRef]
  11. Coombs, G.; Baines, S.; Howden, B.; Swenson, K.; O’Brien, F. Diversity of bacteriophages encoding Panton-Valentine leukocidin in temporally and geographically related Staphylococcus aureus. PLoS ONE 2020, 15, e0228676. [Google Scholar] [CrossRef] [PubMed]
  12. Baba, T.; Takeuchi, F.; Kuroda, M.; Yuzawa, H.; Aoki, K.; Oguchi, A.; Nagai, Y.; Iwama, N.; Asano, K.; Naimi, T.; et al. Genome and virulence determinants of high virulence community-acquired MRSA. Lancet 2002, 359, 1819–1827. [Google Scholar] [CrossRef]
  13. Monecke, S.; Burgold-Voigt, S.; Braun, S.D.; Diezel, C.; Liebler-Tenorio, E.M.; Muller, E.; Nassar, R.; Reinicke, M.; Reissig, A.; Senok, A.; et al. Characterisation of PVL-Positive Staphylococcus argenteus from the United Arab Emirates. Antibiotics 2024, 13, 401. [Google Scholar] [CrossRef] [PubMed]
  14. Jayasinghe, L.; Bayley, H. The leukocidin pore: Evidence for an octamer with four LukF subunits and four LukS subunits alternating around a central axis. Protein Sci. 2005, 14, 2550–2561. [Google Scholar] [CrossRef]
  15. Löffler, B.; Hussain, M.; Grundmeier, M.; Bruck, M.; Holzinger, D.; Varga, G.; Roth, J.; Kahl, B.C.; Proctor, R.A.; Peters, G. Staphylococcus aureus panton-valentine leukocidin is a very potent cytotoxic factor for human neutrophils. PLoS Pathog. 2010, 6, e1000715. [Google Scholar] [CrossRef] [PubMed]
  16. Simpson, V.; Davison, N.; Hudson, L.; Whatmore, A.M. Staphylococcus aureus ST49 infection in red squirrels. Vet. Rec. 2010, 167, 69. [Google Scholar] [CrossRef]
  17. Simpson, V.R.; Davison, N.J.; Kearns, A.M.; Pichon, B.; Hudson, L.O.; Koylass, M.; Blackett, T.; Butler, H.; Rasigade, J.P.; Whatmore, A.M. Association of a lukM-positive clone of Staphylococcus aureus with fatal exudative dermatitis in red squirrels (Sciurus vulgaris). Vet. Microbiol. 2013, 162, 987–991. [Google Scholar] [CrossRef]
  18. Koop, G.; Vrieling, M.; Storisteanu, D.M.; Lok, L.S.; Monie, T.; van Wigcheren, G.; Raisen, C.; Ba, X.; Gleadall, N.; Hadjirin, N.; et al. Identification of LukPQ, a novel, equid-adapted leukocidin of Staphylococcus aureus. Sci. Rep. 2017, 7, 40660. [Google Scholar] [CrossRef]
  19. Monecke, S.; Burgold-Voigt, S.; Feßler, A.T.; Krapf, M.; Loncaric, I.; Liebler-Tenorio, E.M.; Braun, S.D.; Diezel, C.; Müller, E.; Reinicke, M.; et al. Characterisation of Staphylococcus aureus Strains and Their Prophages That Carry Horse-Specific Leukocidin Genes lukP/Q. Toxins 2025, 17, 20. [Google Scholar] [CrossRef]
  20. Monecke, S.; Feßler, A.T.; Burgold-Voigt, S.; Krüger, H.; Mühldorfer, K.; Wibbelt, G.; Liebler-Tenorio, E.M.; Reinicke, M.; Braun, S.D.; Hanke, D.; et al. Staphylococcus aureus isolates from Eurasian Beavers (Castor fiber) carry a novel phage-borne bicomponent leukocidin related to the Panton-Valentine leukocidin. Sci. Rep. 2021, 11, 24394. [Google Scholar] [CrossRef]
  21. Gillet, Y.; Issartel, B.; Vanhems, P.; Fournet, J.C.; Lina, G.; Bes, M.; Vandenesch, F.; Piemont, Y.; Brousse, N.; Floret, D.; et al. Association between Staphylococcus aureus strains carrying gene for Panton-Valentine leukocidin and highly lethal necrotising pneumonia in young immunocompetent patients. Lancet 2002, 359, 753–759. [Google Scholar] [CrossRef] [PubMed]
  22. Monecke, S.; Luedicke, C.; Slickers, P.; Ehricht, R. Molecular epidemiology of Staphylococcus aureus in asymptomatic carriers. Eur. J. Clin. Microbiol. Infect. Dis. 2009, 28, 1159–1165. [Google Scholar] [CrossRef]
  23. Monecke, S.; Slickers, P.; Ellington, M.; Kearns, A.; Ehricht, R. High diversity of Panton-Valentine leucocidin-positive, methicillin-susceptible isolates of Staphylococcus aureus and implications for the evolution of community-associated MRSA. Clin. Microbiol. Infect. 2007, 13, 1157–1164. [Google Scholar] [CrossRef]
  24. Darboe, S.; Dobreniecki, S.; Jarju, S.; Jallow, M.; Mohammed, N.I.; Wathuo, M.; Ceesay, B.; Tweed, S.; Basu Roy, R.; Okomo, U.; et al. Prevalence of Panton-Valentine Leukocidin (PVL) and Antimicrobial Resistance in Community-Acquired Clinical Staphylococcus aureus in an Urban Gambian Hospital: A 11-Year Period Retrospective Pilot Study. Front. Cell. Infect Microbiol. 2019, 9, 170. [Google Scholar] [CrossRef] [PubMed]
  25. Grebe, T.; Rudolf, V.; Gouleu, C.S.; Löffler, B.; Adegnika, A.A.; Shittu, A.O.; Deinhardt-Emmer, S.; Niemann, S.; Schaumburg, F. Neutralization of the Staphylococcus aureus Panton-Valentine leukocidin by African and Caucasian sera. BMC Microbiol. 2022, 22, 219. [Google Scholar] [CrossRef]
  26. Dunne, W.M., Jr. Panton-Valentine leukocidin genes in a laboratory quality control strain of Staphylococcus aureus. J. Clin. Microbiol. 2006, 44, 287. [Google Scholar] [CrossRef] [PubMed]
  27. Kearns, A.M.; Ganner, M.; Holmes, A. The ‘Oxford Staphylococcus’: A note of caution. J. Antimicrob. Chemother. 2006, 58, 480–481. [Google Scholar] [CrossRef]
  28. Frazee, B.W.; Lynn, J.; Charlebois, E.D.; Lambert, L.; Lowery, D.; Perdreau-Remington, F. High prevalence of methicillin-resistant Staphylococcus aureus in emergency department skin and soft tissue infections. Ann. Emerg. Med. 2005, 45, 311–320. [Google Scholar] [CrossRef]
  29. Moran, G.J.; Krishnadasan, A.; Gorwitz, R.J.; Fosheim, G.E.; McDougal, L.K.; Carey, R.B.; Talan, D.A.; Group, E.M.I.N.S. Methicillin-resistant S. aureus infections among patients in the emergency department. N. Engl. J. Med. 2006, 355, 666–674. [Google Scholar] [CrossRef]
  30. Monecke, S.; Baier, V.; Coombs, G.W.; Slickers, P.; Ziegler, A.; Ehricht, R. Genome sequencing and molecular characterisation of Staphylococcus aureus ST772-MRSA-V, “Bengal Bay Clone”. BMC Res. Notes 2013, 6, 1–7. [Google Scholar] [CrossRef]
  31. Steinig, E.J.; Duchene, S.; Robinson, D.A.; Monecke, S.; Yokoyama, M.; Laabei, M.; Slickers, P.; Andersson, P.; Williamson, D.; Kearns, A.; et al. Evolution and Global Transmission of a Multidrug-Resistant, Community-Associated Methicillin-Resistant Staphylococcus aureus Lineage from the Indian Subcontinent. mBio 2019, 10, e01105-19. [Google Scholar] [CrossRef]
  32. Prabhakara, S.; Khedkar, S.; Loganathan, R.M.; Chandana, S.; Gowda, M.; Arakere, G.; Seshasayee, A.S.N. Draft Genome Sequence of Staphylococcus aureus 118 (ST772), a Major Disease Clone from India. J. Bacteriol. 2012, 194, 3727–3728. [Google Scholar] [CrossRef]
  33. D’Souza, N.; Rodrigues, C.; Mehta, A. Molecular characterization of methicillin-resistant Staphylococcus aureus with emergence of epidemic clones of sequence type (ST) 22 and ST 772 in Mumbai, India. J. Clin. Microbiol. 2010, 48, 1806–1811. [Google Scholar] [CrossRef]
  34. Coombs, G.; Pearson, J.; Daley, D.; Robinson, O.; Nimmo, G.; Turnidge, J.D. Staphylococcus aureus Programme 2012 (SAP 2012) Community Survey, MRSA Epidemiology and Typing Report; Australian Group on Antimicrobial Resistance: East Perth, Australia, 2013; Available online: https://agargroup.org.au/wp-content/uploads/2022/09/FED-REPORT-SAP2012-MRSA-Typing-Report-FINAL.pdf (accessed on 26 January 2025).
  35. Coombs, G.W.; Goering, R.V.; Chua, K.Y.; Monecke, S.; Howden, B.P.; Stinear, T.P.; Ehricht, R.; O’Brien, F.G.; Christiansen, K.J. The Molecular Epidemiology of the Highly Virulent ST93 Australian Community Staphylococcus aureus Strain. PLoS ONE 2012, 7, e43037. [Google Scholar] [CrossRef] [PubMed]
  36. Coombs, G.W.; Nimmo, G.R.; Bell, J.M.; Huygens, F.; O’Brien, F.G.; Malkowski, M.J.; Pearson, J.C.; Stephens, A.J.; Giffard, P.M.; Resistance, A.G.A. Genetic diversity among community methicillin-resistant Staphylococcus aureus strains causing outpatient infections in Australia. J. Clin. Microbiol. 2004, 42, 4735–4743. [Google Scholar] [CrossRef]
  37. Coombs, G.W.; Pearson, J.C.; O’Brien, F.G.; Murray, R.J.; Grubb, W.B.; Christiansen, K.J. Methicillin-resistant Staphylococcus aureus clones, Western Australia. Emerg. Infect. Dis. 2006, 12, 241–247. [Google Scholar] [CrossRef] [PubMed]
  38. Chroboczek, T.; Boisset, S.; Rasigade, J.P.; Meugnier, H.; Akpaka, P.E.; Nicholson, A.; Nicolas, M.; Olive, C.; Bes, M.; Vandenesch, F.; et al. Major West Indies MRSA clones in human beings: Do they travel with their hosts? J. Travel Med. 2013, 20, 283–288. [Google Scholar] [CrossRef] [PubMed]
  39. Helgason, K.O.; Jones, M.E.; Edwards, G. Panton-valentine leukocidin-positive Staphylococcus aureus and foreign travel. J. Clin. Microbiol. 2008, 46, 832–833. [Google Scholar] [CrossRef]
  40. Maier, J.; Melzl, H.; Reischl, U.; Drubel, I.; Witte, W.; Lehn, N.; Linde, H. Panton-Valentine leukocidin-positive methicillin-resistant Staphylococcus aureus in Germany associated with travel or foreign family origin. Eur. J. Clin. Microbiol. Infect. Dis. 2005, 24, 637–639. [Google Scholar] [CrossRef]
  41. Nurjadi, D.; Friedrich-Janicke, B.; Schafer, J.; Van Genderen, P.J.; Goorhuis, A.; Perignon, A.; Neumayr, A.; Mueller, A.; Kantele, A.; Schunk, M.; et al. Skin and soft tissue infections in intercontinental travellers and the import of multi-resistant Staphylococcus aureus to Europe. Clin. Microbiol. Infect. 2015, 21, 567.e1–567.e10. [Google Scholar] [CrossRef]
  42. Zanger, P.; Nurjadi, D.; Schleucher, R.; Scherbaum, H.; Wolz, C.; Kremsner, P.G.; Schulte, B. Import and spread of Panton-Valentine Leukocidin-positive Staphylococcus aureus through nasal carriage and skin infections in travelers returning from the tropics and subtropics. Clin. Infect. Dis. 2012, 54, 483–492. [Google Scholar] [CrossRef]
  43. PVL Subgroup of the Steering Group on Healthcare Associated Infection. Guidance on the Diagnosis and Management of PVL-Associated Staphylococcus aureus Infections (PVL-SA) in England; Health Protection Agency: London, UK, 2008. [Google Scholar]
  44. Leistner, R.; Hanitsch, L.G.; Krüger, R.; Lindner, A.K.; Stegemann, M.S.; Nurjadi, D. Skin Infections Due to Panton-Valentine Leukocidin–Producing S. aureus. Dtsch Arztebl. Int. 2022, 119, 775–784. [Google Scholar] [CrossRef] [PubMed]
  45. Merbecks, S.S. Neue bundesweit geltende Meldepflichten: Bedeutung für Sachsen. Ärzteblatt Sachs. 2016, 235–236. Available online: https://www.slaek.de/media/dokumente/ueber-uns/presse/aerzteblatt/archiv/2011-2020/2016/aebl0616.pdf (accessed on 26 January 2025).
  46. Sächsische Infektionsschutz-Meldeverordnung vom 19. Juli 2024 (SächsGVBl. S. 745, Fsn-Nr.: 250-6/3 ). SächsGVBl 2024, 745.
  47. Köhler, G.; Milstein, C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975, 256, 495–497. [Google Scholar] [CrossRef]
  48. Monecke, S.; Slickers, P.; Ehricht, R. Assignment of Staphylococcus aureus isolates to clonal complexes based on microarray analysis and pattern recognition. FEMS Immunol. Med. Microbiol. 2008, 53, 237–251. [Google Scholar] [CrossRef] [PubMed]
  49. Monecke, S.; Coombs, G.; Shore, A.C.; Coleman, D.C.; Akpaka, P.; Borg, M.; Chow, H.; Ip, M.; Jatzwauk, L.; Jonas, D.; et al. A field guide to pandemic, epidemic and sporadic clones of methicillin-resistant Staphylococcus aureus. PLoS ONE 2011, 6, e17936. [Google Scholar] [CrossRef] [PubMed]
  50. Monecke, S.; Gavier-Widen, D.; Mattsson, R.; Rangstrup-Christensen, L.; Lazaris, A.; Coleman, D.C.; Shore, A.C.; Ehricht, R. Detection of mecC-positive Staphylococcus aureus (CC130-MRSA-XI) in diseased European hedgehogs (Erinaceus europaeus) in Sweden. PLoS ONE 2013, 8, e66166. [Google Scholar] [CrossRef]
  51. Stieber, B.; Monecke, S.; Müller, E.; Ehricht, R. Measuring of S. aureus Panton-Valentine leukocidin expression and regulation using protein microarrays. Int. J. Med. Microbiol. 2012, 302, 66–67. [Google Scholar]
  52. Stieber, B.; Sabat, A.; Monecke, S.; Slickers, P.; Akkerboom, V.; Müller, E.; Friedrich, A.W.; Ehricht, R. PVL overexpression due to genomic rearrangements and mutations in the S. aureus reference strain ATCC25923. BMC Res. Notes 2017, 10, 576. [Google Scholar] [CrossRef]
  53. Noda, M.; Kato, I. Purification and crystallization of staphylococcal leukocidin. Methods Enzymol. 1988, 165, 22–32. [Google Scholar] [CrossRef]
  54. Monecke, S.; Akpaka, P.E.; Smith, M.R.; Unakal, C.G.; Thoms Rodriguez, C.A.; Ashraph, K.; Müller, E.; Braun, S.D.; Diezel, C.; Reinicke, M.; et al. Clonal Complexes Distribution of Staphylococcus aureus Isolates from Clinical Samples from the Caribbean Islands. Antibiotics 2023, 12, 1050. [Google Scholar] [CrossRef]
  55. Monecke, S.; Müller, E.; Buechler, J.; Rejman, J.; Stieber, B.; Akpaka, P.E.; Bandt, D.; Burris, R.; Coombs, G.; Hidalgo-Arroyo, G.A.; et al. Rapid detection of Panton-Valentine leukocidin in Staphylococcus aureus cultures by use of a lateral flow assay based on monoclonal antibodies. J. Clin. Microbiol. 2013, 51, 487–495. [Google Scholar] [CrossRef] [PubMed]
  56. Linde, H.; Wagenlehner, F.; Strommenger, B.; Drubel, I.; Tanzer, J.; Reischl, U.; Raab, U.; Holler, C.; Naber, K.G.; Witte, W.; et al. Healthcare-associated outbreaks and community-acquired infections due to MRSA carrying the Panton-Valentine leucocidin gene in southeastern Germany. Eur. J. Clin. Microbiol. Infect. Dis. 2005, 24, 419–422. [Google Scholar] [CrossRef] [PubMed]
  57. Monecke, S.; Jatzwauk, L.; Müller, E.; Nitschke, H.; Pfohl, K.; Slickers, P.; Reissig, A.; Ruppelt-Lorz, A.; Ehricht, R. Diversity of SCCmec elements in Staphylococcus aureus as observed in South-Eastern Germany. PLoS ONE 2016, 11, e0162654. [Google Scholar] [CrossRef]
  58. Monecke, S.; Jatzwauk, L.; Ruppelt, A.; Albrecht, N.; Stieber, B.; Ehricht, R. The rise of the Barnim epidemic strain (CC22-MRSA-IV)—Changes affecting the MRSA population structure in a German university hospital during fourteen years. Int. J. Med. Microbiol. 2013, 303, 84. [Google Scholar]
  59. Monecke, S.; Jatzwauk, L.; Slickers, P.; Ehricht, R. Surveillance of emerging community acquired MRSA: The situation in Dresden/Saxony. Int. J. Med. Microbiol. 2008, 298, 59. [Google Scholar]
  60. Monecke, S.; Jatzwauk, L.; Weber, S.; Slickers, P.; Ehricht, R. DNA microarray-based genotyping of methicillin-resistant Staphylococcus aureus strains from Eastern Saxony. Clin. Microbiol. Infect. 2008, 14, 534–545. [Google Scholar] [CrossRef] [PubMed]
  61. Egyir, B.; Guardabassi, L.; Sorum, M.; Nielsen, S.S.; Kolekang, A.; Frimpong, E.; Addo, K.K.; Newman, M.J.; Larsen, A.R. Molecular epidemiology and antimicrobial susceptibility of clinical Staphylococcus aureus from healthcare institutions in Ghana. PLoS ONE 2014, 9, e89716. [Google Scholar] [CrossRef]
  62. Ruimy, R.; Maiga, A.; Armand-Lefevre, L.; Maiga, I.; Diallo, A.; Koumare, A.K.; Ouattara, K.; Soumare, S.; Gaillard, K.; Lucet, J.C.; et al. The carriage population of Staphylococcus aureus from Mali is composed of a combination of pandemic clones and the divergent Panton-Valentine leukocidin-positive genotype ST152. J. Bacteriol. 2008, 190, 3962–3968. [Google Scholar] [CrossRef]
  63. Shittu, A.; Oyedara, O.; Abegunrin, F.; Okon, K.; Raji, A.; Taiwo, S.; Ogunsola, F.; Onyedibe, K.; Elisha, G. Characterization of methicillin-susceptible and -resistant staphylococci in the clinical setting: A multicentre study in Nigeria. BMC Infect. Dis. 2012, 12, 286. [Google Scholar] [CrossRef]
  64. Shittu, A.O.; Okon, K.; Adesida, S.; Oyedara, O.; Witte, W.; Strommenger, B.; Layer, F.; Nubel, U. Antibiotic resistance and molecular epidemiology of Staphylococcus aureus in Nigeria. BMC Microbiol. 2011, 11, 92. [Google Scholar] [CrossRef] [PubMed]
  65. Breurec, S.; Fall, C.; Pouillot, R.; Boisier, P.; Brisse, S.; Diene-Sarr, F.; Djibo, S.; Etienne, J.; Fonkoua, M.C.; Perrier-Gros-Claude, J.D.; et al. Epidemiology of methicillin-susceptible Staphylococcus aureus lineages in five major African towns: High prevalence of Panton-Valentine leukocidin genes. Clin. Microbiol. Infect. 2011, 17, 633–639. [Google Scholar] [CrossRef] [PubMed]
  66. Strauß, L.; Ruffing, U.; Abdulla, S.; Alabi, A.; Akulenko, R.; Garrine, M.; Germann, A.; Grobusch, M.P.; Helms, V.; Herrmann, M.; et al. Detecting Staphylococcus aureus Virulence and Resistance Genes: A Comparison of Whole-Genome Sequencing and DNA Microarray Technology. J. Clin. Microbiol. 2016, 54, 1008–1016. [Google Scholar] [CrossRef]
  67. Monecke, S.; Bedewy, A.K.; Müller, E.; Braun, S.D.; Diezel, C.; Elsheredy, A.; Kader, O.; Reinicke, M.; Ghazal, A.; Rezk, S.; et al. Characterisation of Methicillin-Resistant Staphylococcus aureus from Alexandria, Egypt. Antibiotics 2023, 12, 78. [Google Scholar] [CrossRef]
  68. Montelongo, C.; Mores, C.R.; Putonti, C.; Wolfe, A.J.; Abouelfetouh, A.; Mkrtchyan, H.V. Whole-Genome Sequencing of Staphylococcus aureus and Staphylococcus haemolyticus Clinical Isolates from Egypt. Microbiol. Spectr. 2022, 10, e02413-21. [Google Scholar] [CrossRef]
  69. Goudarzi, M.; Goudarzi, H.; Sá Figueiredo, A.M.; Udo, E.E.; Fazeli, M.; Asadzadeh, M.; Seyedjavadi, S.S. Molecular Characterization of Methicillin Resistant Staphylococcus aureus Strains Isolated from Intensive Care Units in Iran: ST22-SCCmec IV/t790 Emerges as the Major Clone. PLoS ONE 2016, 11, e0155529. [Google Scholar] [CrossRef]
  70. Senok, A.; Monecke, S.; Nassar, R.; Celiloglu, H.; Thyagarajan, S.; Müller, E.; Ehricht, R. Lateral Flow Immunoassay for the Detection of Panton-Valentine Leukocidin in Staphylococcus aureus from skin and soft tissue infections in the United Arab Emirates. Front. Microbiol. 2021, 11, 754523. [Google Scholar] [CrossRef]
  71. Zhao, H.; Wu, X.; Wang, B.; Shen, L.; Rao, L.; Wang, X.; Zhang, J.; Xiao, Y.; Xu, Y.; Yu, J.; et al. Phenotypic and genomic analysis of the hypervirulent ST22 methicillin-resistant Staphylococcus aureus in China. mSystems 2023, 8, e0124222. [Google Scholar] [CrossRef] [PubMed]
  72. Roberts, M.C.; Joshi, P.R.; Greninger, A.L.; Melendez, D.; Paudel, S.; Acharya, M.; Bimali, N.K.; Koju, N.P.; No, D.; Chalise, M.; et al. The human clone ST22 SCCmec IV methicillin-resistant Staphylococcus aureus isolated from swine herds and wild primates in Nepal: Is man the common source? FEMS Microbiol. Ecol. 2018, 94, fiy052. [Google Scholar] [CrossRef]
  73. Reyes, J.; Rincon, S.; Diaz, L.; Panesso, D.; Contreras, G.A.; Zurita, J.; Carrillo, C.; Rizzi, A.; Guzman, M.; Adachi, J.; et al. Dissemination of methicillin-resistant Staphylococcus aureus USA300 sequence type 8 lineage in Latin America. Clin. Infect. Dis. 2009, 49, 1861–1867. [Google Scholar] [CrossRef]
  74. Planet, P.J.; Diaz, L.; Rios, R.; Arias, C.A. Global Spread of the Community-Associated Methicillin-Resistant Staphylococcus aureus USA300 Latin American Variant. J. Infect. Dis. 2016, 214, 1609–1610. [Google Scholar] [CrossRef] [PubMed]
  75. Adhikari, R.P.; Cook, G.M.; Lamont, I.; Lang, S.; Heffernan, H.; Smith, J.M. Phenotypic and molecular characterization of community occurring, Western Samoan phage pattern methicillin-resistant Staphylococcus aureus. J. Antimicrob. Chemother. 2002, 50, 825–831. [Google Scholar] [CrossRef] [PubMed]
  76. Smith, J.M.; Cook, G.M. A decade of community MRSA in New Zealand. Epidemiol. Infect. 2005, 133, 899–904. [Google Scholar] [CrossRef] [PubMed]
  77. Aloba, B.K.; Kinnevey, P.M.; Monecke, S.; Brennan, G.I.; O’Connell, B.; Blomfeldt, A.; McManus, B.A.; Schneider-Brachert, W.; Tkadlec, J.; Ehricht, R.; et al. An emerging Panton-Valentine leukocidin-positive CC5-meticillin-resistant Staphylococcus aureus-IVc clone recovered from hospital and community settings over a 17-year period from 12 countries investigated by whole-genome sequencing. J. Hosp. Infect. 2023, 132, 8–19. [Google Scholar] [CrossRef]
  78. McManus, B.A.; Aloba, B.K.; Earls, M.R.; Brennan, G.I.; O’Connell, B.; Monecke, S.; Ehricht, R.; Shore, A.C.; Coleman, D.C. Multiple distinct outbreaks of Panton-Valentine leukocidin (PVL)-positive community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) in Ireland investigated by whole-genome sequencing. J. Hosp. Infect. 2020, 108, 72–80. [Google Scholar] [CrossRef]
  79. Witte, W.; Enright, M.; Schmitz, F.J.; Cuny, C.; Braulke, C.; Heuck, D. Characteristics of a new epidemic MRSA in Germany ancestral to United Kingdom EMRSA 15. Int. J. Med. Microbiol. 2001, 290, 677–682. [Google Scholar] [CrossRef]
  80. Earls, M.R.; Kinnevey, P.M.; Brennan, G.I.; Lazaris, A.; Skally, M.; O’Connell, B.; Humphreys, H.; Shore, A.C.; Coleman, D.C. The recent emergence in hospitals of multidrug-resistant community-associated sequence type 1 and spa type t127 methicillin-resistant Staphylococcus aureus investigated by whole-genome sequencing: Implications for screening. PLoS ONE 2017, 12, e0175542. [Google Scholar] [CrossRef]
  81. Monecke, S.; Konig, E.; Earls, M.R.; Leitner, E.; Müller, E.; Wagner, G.E.; Poitz, D.M.; Jatzwauk, L.; Vremera, T.; Dorneanu, O.S.; et al. An epidemic CC1-MRSA-IV clone yields false-negative test results in molecular MRSA identification assays: A note of caution, Austria, Germany, Ireland, 2020. Eurosurveillance 2020, 25, 2000929. [Google Scholar] [CrossRef]
  82. Earls, M.R.; Steinig, E.J.; Monecke, S.; Samaniego Castruita, J.A.; Simbeck, A.; Schneider-Brachert, W.; Vremerǎ, T.; Dorneanu, O.S.; Loncaric, I.; Bes, M.; et al. Exploring the evolution and epidemiology of European CC1-MRSA-IV: Tracking a multidrug-resistant community-associated meticillin-resistant Staphylococcus aureus clone. Microb. Genom. 2021, 7, 000601. [Google Scholar] [CrossRef]
  83. Scicluna, E.A.; Shore, A.C.; Thurmer, A.; Ehricht, R.; Slickers, P.; Borg, M.A.; Coleman, D.C.; Monecke, S. Characterisation of MRSA from Malta and the description of a Maltese epidemic MRSA strain. Eur. J. Clin. Microbiol. Infect. Dis. 2010, 29, 163–170. [Google Scholar] [CrossRef]
  84. Monecke, S.; Skakni, L.; Hasan, R.; Ruppelt, A.; Ghazal, S.S.; Hakawi, A.; Slickers, P.; Ehricht, R. Characterisation of MRSA strains isolated from patients in a hospital in Riyadh, Kingdom of Saudi Arabia. BMC Microbiol. 2012, 12, 146. [Google Scholar] [CrossRef] [PubMed]
  85. Senok, A.; Ehricht, R.; Monecke, S.; Al-Saedan, R.; Somily, A. Molecular characterization of methicillin-resistant Staphylococcus aureus in nosocomial infections in a tertiary-care facility: Emergence of new clonal complexes in Saudi Arabia. New Microbes New Infect. 2016, 14, 13–18. [Google Scholar] [CrossRef] [PubMed]
  86. Senok, A.; Nassar, R.; Kaklamanos, E.G.; Belhoul, K.; Abu Fanas, S.; Nassar, M.; Azar, A.J.; Muller, E.; Reissig, A.; Gawlik, D.; et al. Molecular characterization of Staphylococcus aureus isolates associated with nasal colonization and environmental contamination in academic dental clinics. Microb. Drug Resist. 2020, 26, 661–669. [Google Scholar] [CrossRef]
  87. Monecke, S.; Müller, E.; Braun, S.D.; Armengol-Porta, M.; Bes, M.; Boswihi, S.; El-Ashker, M.; Engelmann, I.; Gawlik, D.; Gwida, M.; et al. Characterisation of S. aureus/MRSA CC1153 and review of mobile genetic elements carrying the fusidic acid resistance gene fusC. Sci. Rep. 2021, 11, 8128. [Google Scholar] [CrossRef]
  88. Senok, A.; Somily, A.M.; Nassar, R.; Garaween, G.; Kim Sing, G.; Müller, E.; Reissig, A.; Gawlik, D.; Ehricht, R.; Monecke, S. Emergence of novel methicillin-resistant Staphylococcus aureus strains in a tertiary care facility in Riyadh, Saudi Arabia. Infect. Drug Resist. 2019, 12, 2739–2746. [Google Scholar] [CrossRef] [PubMed]
  89. Senok, A.; Somily, A.; Raji, M.; Garaween, G.; Kabil, M.; Shibl, A.; Monecke, S.; Ehricht, R. Genotyping of Staphylococcus aureus associated with nasal colonization among healthcare workers using DNA microarray. J. Infect. Dev. Ctries 2018, 12, 321–325. [Google Scholar] [CrossRef]
  90. Senok, A.; Nassar, R.; Celiloglu, H.; Nabi, A.; Alfaresi, M.; Weber, S.; Rizvi, I.; Müller, E.; Reissig, A.; Gawlik, D.; et al. Genotyping of methicillin resistant Staphylococcus aureus from the United Arab Emirates. Sci. Rep. 2020, 10, 18551. [Google Scholar] [CrossRef]
  91. Boswihi, S.S.; Udo, E.E.; Monecke, S.; Mathew, B.; Noronha, B.; Verghese, T.; Tappa, S.B. Emerging variants of methicillin-resistant Staphylococcus aureus genotypes in Kuwait hospitals. PLoS ONE 2018, 13, e0195933. [Google Scholar] [CrossRef]
  92. Müller, E.; Monecke, S.; Armengol Porta, M.; Narvaez Encalada, M.V.; Schröttner, P.; Schwede, I.; Söffing, H.-H.; Thürmer, A.; Ehricht, R. Characterisation of PVL-positive Staphylococcus aureus isolates from Saxony and Brandenburg. In Proceedings of the Annual Conference of the German Society for Microbiology and Hygiene, Würzburg, Germany, 2–5 June 2024; p. 132. [Google Scholar]
Pathogens 14 00238 g001
Figure 1. Comparison to the results of the German branch of the previous study [55].
Figure 1. Comparison to the results of the German branch of the previous study [55].
Pathogens 14 00238 g001
Table 1. Strain affiliations, PVL- und mecA status of study isolates.
Table 1. Strain affiliations, PVL- und mecA status of study isolates.
CategoryCCStrainNPercent
PVL-negative MSSA3542.2%
CC5CC5-MSSA11.2%
CC7CC7-MSSA78.4%
CC10CC10-MSSA22.4%
CC15CC15-MSSA44.8%
CC15 (ST199)-MSSA11.2%
CC15 (ST582)-MSSA22.4%
CC22CC22-MSSA44.8%
CC22-MSSA-[ccrAB4]22.4%
CC25CC25-MSSA11.2%
CC30CC30-MSSA44.8%
CC45CC45-MSSA44.8%
CC398CC398-MSSA22.4%
ST2867ST2867-MSSA11.2%
PVL-negative MRSA910.8%
CC1CC1-MRSA-IV (aphA3/sat-positive), “European CC1-MRSA-IV strain”11.2%
CC1-MRSA-[IV+fus+ccrAB1], “WA MRSA-1/45”11.2%
CC5CC5-MRSA-[IV+fus+ccrAB], “Maltese Clone”33.6%
CC5-MRSA-[VI+fus]11.2%
CC22CC22-MRSA-IV, “UK-EMRSA-15/Barnim EMRSA”11.2%
CC22-MRSA-[V/VT+fus]11.2%
CC59CC59-MRSA-[V/VT+fus]11.2%
PVL-positive MSSA1821.7%
CC5CC5-MSSA (PVL+)22.4%
CC6CC6-MSSA (PVL+)11.2%
CC15CC15-MSSA (PVL+)11.2%
CC30CC30-MSSA (PVL+)22.4%
CC88CC88-MSSA (PVL+)11.2%
CC121CC121-MSSA (PVL+)44.8%
CC152CC152-MSSA (PVL+)67.2%
CC772CC772-MSSA (PVL+)11.2%
PVL-positive MRSA2125.3%
CC1CC1-MRSA-[V/VT+fus+ccrAB1] (PVL+)11.2%
CC5CC5-MRSA-IV (sed/j/r+, PVL+), “Sri Lanka Clone”44.8%
CC8CC8-MRSA-[IV+Hg] (PVL+), “Spanish/Latin American “USA300”44.8%
CC8-MRSA-[IV+ACME] (PVL+), „USA300“11.2%
CC22CC22-MRSA-IV (PVL+/tst+)22.4%
CC30CC30-MRSA-IV (PVL+), “WSPP/Southwest Pacific Clone” 11.2%
CC88CC88-MRSA-IV (PVL+)11.2%
CC152CC152-MRSA-IV (PVL+)11.2%
CC152-MRSA-[V/VT+fus] (PVL+)11.2%
CC398CC398-MRSA-V/VT (PVL+)56.0%
Table 2. Resistance genes in study isolates. Percentages in brackets refer to the respective group (PVL-negative MSSA and MRSA, PVL-positive MSSA and MRSA).
Table 2. Resistance genes in study isolates. Percentages in brackets refer to the respective group (PVL-negative MSSA and MRSA, PVL-positive MSSA and MRSA).
GeneExplanation/Corresponding Resistance PhenotypeTotal (n = 83)PVL-Negat. MSSA (n = 35)PVL-Negat. MRSA (n = 9)PVL-Posit. MSSA (n = 18)PVL-Posit. MRSA (n = 23)
mecAMRSA, methicillin/β-lactam 30 (36.1%)09 (100%)021 (100%)
mecCMRSA, methicillin/β-lactam00000
blaZβ-lactamase/penicillin 65 (78.3%)23 (65.7%)9 (100%)13 (72.2%)20 (95.2%)
fusCFusidic acid (SCC-borne)9 (10.8%)07 (77.8%)02 (9.5%)
far1Fusidic acid (plasmid-borne)00000
erm(A)Macrolides, clindamycin5 (6%)0005 (23.8%)
erm(B)Macrolides, clindamycin2 (2.4%)1 (2.9%)001 (4.8%)
erm(C)Macrolides, clindamycin7 (8.4%)02 (22.2%)05 (23.8%)
linA/lnu(A)Lincosamides, clindamycin00000
msr(A)Macrolides4 (4.8%)1 (2.9%)01 (5.6%)2 (9.5%)
mef(A)Macrolides00000
mph(C)Macrolides3 (3.6%)001 (5.6%)2 (9.5%)
aacA-aphDGentamicin, tobramycin7 (8.4%)2 (5.7%)005 (23.8%)
aadDTobramycin3 (3.6%)2 (5.7%)001 (4.8%)
aphA3Kanamycin5 (6%)01 (11.1%)1 (5.6%)3 (14.3%)
dfrATrimethoprim5 (6%)01 (11.1%)04 (19%)
mupAMupirocin (high level)2 (2.4%)1 (2.9%)001 (4.8%)
tet(K)Tetracyclines13 (15.7%)01 (11.1%)2 (11.1%)10 (47.6%)
tet(M)Tetracyclines2 (2.4%)01 (11.1%)1 (5.6%)0
cat (all variants) *Chloramphenicol4 (4.8%)1 (2.9%)02 (11.1%)1 (4.8%)
cfr“PhlopsA” resistance phenotype **00000
fexAChloramphenicol/florfenicol1 (1.2%)01 (11.1%)00
vanAVancomycin, teicoplanin00000
qacA/BBiozide resistance00000
qacC=smrBiozide resistance3 (3.6%)1 (2.9%)01 (5.6%)1 (4.8%)
* For details, see Supplemental file S1; ** “PhlopsA” = Phenicols, Lincosamides, Oxazolidinones, Pleuromutilins, and Streptogramin A.
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Müller, E.; Monecke, S.; Armengol Porta, M.; Narvaez Encalada, M.V.; Reissig, A.; Rüttiger, L.; Schröttner, P.; Schwede, I.; Söffing, H.-H.; Thürmer, A.; et al. Rapid Detection of Panton–Valentine Leukocidin Production in Clinical Isolates of Staphylococcus aureus from Saxony and Brandenburg and Their Molecular Characterisation. Pathogens 2025, 14, 238. https://doi.org/10.3390/pathogens14030238

AMA Style

Müller E, Monecke S, Armengol Porta M, Narvaez Encalada MV, Reissig A, Rüttiger L, Schröttner P, Schwede I, Söffing H-H, Thürmer A, et al. Rapid Detection of Panton–Valentine Leukocidin Production in Clinical Isolates of Staphylococcus aureus from Saxony and Brandenburg and Their Molecular Characterisation. Pathogens. 2025; 14(3):238. https://doi.org/10.3390/pathogens14030238

Chicago/Turabian Style

Müller, Elke, Stefan Monecke, Marc Armengol Porta, Marco Vinicio Narvaez Encalada, Annett Reissig, Lukas Rüttiger, Percy Schröttner, Ilona Schwede, Hans-Herman Söffing, Alexander Thürmer, and et al. 2025. "Rapid Detection of Panton–Valentine Leukocidin Production in Clinical Isolates of Staphylococcus aureus from Saxony and Brandenburg and Their Molecular Characterisation" Pathogens 14, no. 3: 238. https://doi.org/10.3390/pathogens14030238

APA Style

Müller, E., Monecke, S., Armengol Porta, M., Narvaez Encalada, M. V., Reissig, A., Rüttiger, L., Schröttner, P., Schwede, I., Söffing, H.-H., Thürmer, A., & Ehricht, R. (2025). Rapid Detection of Panton–Valentine Leukocidin Production in Clinical Isolates of Staphylococcus aureus from Saxony and Brandenburg and Their Molecular Characterisation. Pathogens, 14(3), 238. https://doi.org/10.3390/pathogens14030238

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop