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Diagnostics 2013, 3(2), 222-231; doi:10.3390/diagnostics3020222

Diagnosis of Upper and Lower Respiratory Tract Bacterial Infections with the Use of Multiplex PCR Assays

1 National Meningitis Reference Laboratory, National School of Public Health, 196, Alexandras Avenue, Athens 11521, Greece 2 School of Health Sciences, Department of Biochemistry and Biotechnology, University of Thessaly, Ploutonos 26 & Aiolou str, Larissa 41221, Greece 3 Second Department of Pediatrics, University of Athens School of Medicine, P. and A. Kyriakou Children’s Hospital, Thevon & Levadeias str, Athens 11527, Greece 4 Penteli's Children's Hospital, Microbiology Laboratory, 8 Ippokratous str., Penteli, Attiki 15236, Greece 5 "Evaggelismos" General Hospital of Athens, Microbiology Laboratory, 45–47 Ipsilantou Str, Athens 10676, Greece
* Author to whom correspondence should be addressed.
Received: 1 February 2013 / Revised: 8 March 2013 / Accepted: 21 March 2013 / Published: 26 March 2013
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The investigation of respiratory infections by molecular techniques provides important information about the epidemiology of respiratory disease, especially during the post-vaccination era. The objective of the present study was the detection of bacterial pathogens directly in clinical samples from patients with upper and lower respiratory tract infections using multiplex polymerase chain reaction (PCR) assays developed in our laboratory. Clinical samples taken over a three-year period (2007–2009) and obtained from 349 patients (adults (n = 66); children (n = 283)) with signs and symptoms of certain upper or lower respiratory tract infections, consisted of: bronchoalveolar lavages (BAL, n = 83), pleural fluids (n = 29), and middle-ear aspirates (n = 237). Overall, 212 samples (61%) were confirmed by culture and/or PCR. Among the positive samples, Streptococcus pneumoniae (mainly serotype 3) was predominant (104/212; 49.0%), followed by non-typable Haemophilus influenzae (NTHi) 59/212; 27.8%) and Streptococcus pyogenes (47/212; 22%). Haemophilus influenzae type b was detected in only three samples. The underlying microbiology of respiratory infections is gradually changing in response to various selective pressures, such as vaccine use and antibiotic consumption. The application of multiplex PCR (mPCR) assays is particularly useful since it successfully identified the microorganisms implicated in acute otitis media or lower respiratory tract infections in nearly 75% of patients with a positive result compared to conventional cultures. Non-culture identification of the implicated pneumococcal serotypes is also an important issue for monitoring pneumococcal infections in the era of conjugate pneumococcal vaccines.
Keywords: ear aspirates; pleural fluids; molecular diagnosis ear aspirates; pleural fluids; molecular diagnosis
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Xirogianni, A.; Tsolia, M.; Voyiatzi, A.; Sioumala, M.; Makri, A.; Argyropoulou, A.; Paniara, O.; Markoulatos, P.; Kourea-Kremastinou, J.; Tzanakaki, G. Diagnosis of Upper and Lower Respiratory Tract Bacterial Infections with the Use of Multiplex PCR Assays. Diagnostics 2013, 3, 222-231.

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