|1||2020||Corman et al. (Germany) ||rRT-PCR; First-line screening tool: E gene assay; Confirmatory testing: RdRp gene assay.||Respiratory samples were obtained during 2019 from patients hospitalised at Charité medical Centre.|
Additional samples were selected from biobanks at the Rijksinstituut voor Volksgezondheid en Milieu), Bilthoven, at Erasmus University Medical Center, Rotterdam, at Public Health England, London, and at the University of Hong Kong.
|Preliminary finding with SARS-CoV strain Frankfurt-1 virions grown on Vero cells, E gene and RdRp gene assays produced the best result (5.2 and 3.8 copies per reaction at 95% detection probability, respectively)|
In vitro transcript RNA identical to 2019-nCoV target sequence, 3.9 copies per reaction for the E gene assay and 3.6 copies per reaction for the RdRp assay which were close to the 95% hit rate of 2.9 copies per reaction
When tested for cross-reaction with other coronavirus, there was no reactivity with the assays.
|2||2016||Kim et al. (Korea) ||Six different commercial MERS-CoV RNA detection kits based on rRT-PCR:|
(i) PowerChek (Kogene Biotech, Korea); (ii) DiaPlexQ (SolGent, Korea); (iii) Anyplex (Seegene, Korea)
Screening: envelope gene (upE)
(iv) AccuPower (Bioneer, Korea) (v) LightMix (Roche Molecular Diagnostics, Switzerland) (vi) UltraFast kits (Nanobiosys, Korea)
detect upE and ORF1a simultaneously
|28 nasopharyngeal swabs that were positive for other respiratory viruses were used for specificity and|
18 lower respiratory specimens for clinical sensitivity.
|All six kits correctly identified 8 of 8 (100%) positive clinical specimens. However, based on the findings from the high inhibition panel, PowerChek and AccuPower were the least sensitive to the presence of PCR inhibition.|||
|3||2014||Shirato et al. (Japan) ||Loopamp RNA Amplification Kit (RT-LAMP; Eiken, Tokyo, Japan)||laboratory isolates MERS-CoV diluted with medium containing pharyngeal swabs obtained from healthy adults||Real-time RT-PCR was able to detect at level as low as 1.6 copies of MERS-CoV RNA while RT-LAMP was also able to detect viral RNA at levels as low as 0.7 copies, showing equivalence with the RT-PCR assay.|
There was no cross reactivity with other respiratory viruses, for RT-LAMP
|4||2017||Go et al. (Korea) ||RT-iiPCR|
Target upE and ORF1a gene
|55 sequential sputum samples collected from 12 patients infected with MERS-CoV were obtained from the Chungnam National University Hospital.||Overall agreement between RT-iiPCR assays and reference RT-qPCR assays were 98.06% (95% CI, 94.43–100%) and 99.03% (95% CI, 95.88–100%) for ORF1a and upE assays, respectively.|||
|5||2015||Hashemzadeh et al. (Iran) ||A onestep rRT-PCR assay, based on specific TaqMan probes||UpE and ORF1b was synthesized due to the difficulty in acquiring patient sample||The sensitivity obtained for upE was fewer than ten copies of RNA template per reaction and for ORF1b was 50 or fewer copies per reaction.|||
|SARS-CoV|| || || |
|6||2005||Lau et al. (Hong Kong) ||Real time qRT-PCR; Antibody-based capture ELISA||40 SARS patients hospitalized in HK between March–May 2003||Sensitivities for qRT-PCR (80% for fecal samples and 25% for urine samples) were higher than those of the polyclonal (50% and 5%) and monoclonal (35% and 8%) antibody-based nucleocapsid antigen capture enzyme-linked immunosorbent assays.|||
|7||2003||Lau et al. (Hong Kong) ||Enhanced real-time fluorescent PCR ||80 suspected or probable SARS cases between 1–3 April 2003||The limit of detection of the enhanced real-time PCR method was 102-fold higher than the standard real-time PCR assay and 107-fold higher than conventional PCR methods|
In the clinical aspect, the enhanced real-time PCR method was able to detect 6 cases of SARS-CoV positive samples that were not confirmed by any other assay
|8||2004||Jiang et al. (Taiwan) ||Quantitative, real-time, nested polymerase chain reaction|
|46 patients with suspected or reported SARS April through May 2003 in Taiwan||The single round PCR yielded a minor amplification signal. Nested PCR produce signal without apparent background. The single round RT-PCR detected 15 of 46 positive cases, while the nest real-time PCR detected 17 of 46 cases.|||
|9||2004||Wu et al. (Taiwan) ||Neutralization test,|
(ELISA), (IFA), and (ICT)
|537 probable SARS patients in Taiwan||With the neutralization test as a reference method, the sensitivity, specificity, positive predictive value, and negative predictive value of the test were|
ELISA: 98.2%, 98.7%, 98.7%, and 98.4%
IFA: 99.1%, 87.8%, 88.1% and 99.1%
ICT: 33.6%, 98.2%, 95.7%, and 56.1%
RT-PCR: 52.2%, 78.7%, 74.5%, and 58.1%,
|10||2003||He et al. (Singapore) ||Western blot assay with N195 protein||274 clinical sera which were collected|
from patients suffering from probable or suspected SARS, dengue fever, autoimmune diseases
|The specificity and sensitivity of this test were 98.3 and 90.9% and 40 of 44 clinical SARS samples were positive. |||
|11||2003||Poon et al. (Hong Kong) ||Real time quantitative RT-PCR modified RNA extraction method|
1b region of SARS-CoVCoV
|50 NPA samples collected from days 1–3 of disease onset from SARS patients in whom SARS CoV infections was subsequently serologically confirmed ||From the 50 NPA specimens collected during the first 3 days of illness, the first-generation RT-PCR assay identified 22% positive sample, modification in the RNA extraction method identified 44% positive samples and the combination of the modified RNA extraction method and real-time quantitative PCR technology, identified 80% positive sample.|||
|12||2004||Guan et al. (Hong Kong) ||Specific ELISA Genelabs Diagnostics Pte Ltd. utilizing two recombinant proteins (Gst-N and Gst-U274)||227 clinical serum specimens collected from SARS patients in Hong Kong between 18 March–24 May 2003 and 385 samples from healthy donors. ||For the ELISA, the overall sensitivity was 71.8% and specificity was 99.5%|
For the immunocromatographic test, overall rate of detection of SARS-associated specimens by the rapid test was 70.5% and its specificity was 97.7%.
|13||2003||Hui et al. (Hong Kong) ||RT-PCR||Clinical samples, NPAs (n = 131) and stool specimens (n = 5), provided by the Department of Microbiology, The University of Hong Kong. ||PCR amplifying the N gene gave an average of a 26.0% (6.3 to 60.0%) stronger intensity signal than that for the 1b gene additional sensitive molecular marker for the diagnosis of the SARS coronavirus|||
|14||2004||Mahony et al. (Canada) ||Seven RT-PCR assays include (i) a nested assay with BNI outer and inner primers with polB gene; (ii) a two-step, non-nested assay with the BNI outer primers with polB gene; (iii) a two-step, non-nested RT-PCR assay with Cor-p-F2 and Cor-p-R1 with polB gene; (iv) a one-step RT-PCR, with BNI outer primers and polB gene with SYBR Green detection; (v) a two-step assay amplifying and nucleocapsid gene with SYBR Green detection; (vi) a one-step assay with the same nucleocapsid primers and gene with TaqMan probe and (vii) a commercial RealArt HPA CoV RT-PCR assay (Artus).||68 specimens, including 17 respiratory tract specimens (nasopharyngeal or throat swabs), 29 urine samples, and 22 stool samples, were collected between March–April of 2003 from hospitalized patients with a probable or suspected diagnosis of SARS at Sunnybrook and Women’s College Health Sciences Centre during the Toronto outbreak of SARS.||There is no significant difference in the sensitivity and specificity for the 7 assays|
Assay 1: 94%; 100%
Assay 2: 100%; 94%
Assay 3: 94%; 100%
Assay 4: 100%; 96%
Assay 5: 94%; 100%
Assay 6: 83%; 100%
Assay 7: 94%; 100%
|15.||2004||Liu et al. (Taiwan)||Indirect IFA||Throat wash samples from 17 confirmed SARS adult patients, and 10 healthy controls admitted to the emergency department of the National Taiwan University Hospital between 16 April–1 May 2003.||SARS-CoV was detected in 11 of 17 (65%) samples from SARS patients from day 2 to day 9 of the illness but in none of the 10 samples from healthy controls|||
|16.||2004||Lin et al. (Taiwan)||SARS-CoV real-time PCR assay with a TaqMan minor groove binder probe developed by Applied Biosystems (Foster City, CA, USA).||228 samples (137 sputum, 53 throat swabs or throat wash, 17 NPS, 19 stool specimen, 2 pleural fluid, 2 urine and 1 serum sample) from 151 patients with atypical|
pneumonia or symptoms mimicking SARS between 30 April–26 June 26 were recruited,
In total, from 151 patients were tested.
|• The real time PCR has a threshold sensitivity of 10 genome equivalents per reaction and it has a good reproducibility with the inter-assay coefficients of variation of 1.73 to 2.72%.|
• 13 specimens from 6 patients were positive with viral load range from 362 to 36,240,000 genome equivalents/mL.
The real-time RT-PCR reaction was more sensitive than the nested PCR reaction, as the detection limit for the nested PCR reaction was about 103 genome equivalents in the standard cDNA control.