key: cord-1049927-6qsb7ry0 authors: Garg, Atul; Ghoshal, Ujjala; Patel, Sangram S.; Singh, D. V.; Arya, Akshay K.; Vasanth, Shruthi; Pandey, Ankita; Srivastava, Nikki title: Evaluation of seven commercial RT‐PCR kits for COVID‐19 testing in pooled clinical specimens date: 2020-12-17 journal: J Med Virol DOI: 10.1002/jmv.26691 sha: 308bc8f00bac98874427bb7d5a0ecd7c4a4361b7 doc_id: 1049927 cord_uid: 6qsb7ry0 There are more than 350 real‐time polymerase chain reaction (RT‐PCR) coronavirus disease‐2019 (COVID‐19) testing kits commercially available but these kits have not been evaluated for pooled sample testing. Thus, this study was planned to compare and evaluate seven commercially available kits for pooled samples testing. Diagnostic accuracy of (1) TRUPCR SARS‐CoV‐2 Kit (Black Bio), (2) TaqPath RT‐PCR COVID‐19 Kit (Thermo Fisher Scientific), (3) Allplex 2019‐nCOV Assay (Seegene), (4) Patho detect COVID‐19 PCR kit (My Lab), (5) LabGun COVID‐19 RT‐PCR Kit (Lab Genomics, Korea), (6) Fosun COVID‐19 RT‐PCR detection kit (Fosun Ltd.), (7) Real‐time Fluorescent RT‐PCR kit for SARS CoV‐2 (BGI) was evaluated on precharacterised 40 positive and 10 negative COVID‐19 sample pools. All seven kits detected all sample pools with low C (t) values (<30); while testing weak positive pooled samples with high C (t) value (>30); the TRUPCR Kit, TaqPath Kit, Allplex Assay, and BGI RT‐PCR kit showed 100% sensitivity, specificity, and accuracy. However, the Fosun kit, LabGun Kit, and Patho detect kit could detect only 90%, 85%, and 75% of weakly positive samples, respectively. We conclude that all seven commercially available RT‐PCR kits included in this study can be used for routine molecular diagnosis of COVID‐19. However, regarding performing pooled sample testing, it might be advisable to use those kits that performed best regarding positive identification in samples' pool, that is TRUPCR SARS‐CoV‐2 Kit, TaqPath RT‐PCR COVID‐19 Kit, Allplex 2019‐nCOV Assay, and BGI Real‐time RT‐PCR kit for detecting SARS CoV‐2. There are more than 350 Conventional RT-PCR COVID-19 testing kits available commercially, of which 29 kits have been approved by the United States food and drug administration (US-FDA). 6 RT-PCR is an expensive test and requires a well equipped molecular laboratory with trained manpower. Many countries are experiencing acute shortages of diagnostic kits and manufacturers of molecular testing kits and consumables are also struggling to keep with the demand. It has become important to come up with novel ideas to conserve the reagents used for molecular tests. However, at the same time, the disease is new it is important to validate modifications to the testing protocol before universal adoption. Several researchers are advocating that it is time to reintroduce the Dorfman theory 7 of sample pooling in the era of molecular testing. 8, 9 In a recent study from the University of Nebraska Medical Centre, Omaha, the authors have used a web-based application and determined the most efficient pool size to be five samples when the incidence rate of SARS-CoV-2 infection is 10% or less and concluded that group testing will result in saving of reagents and increase in the testing capability of at least 69%. 10 Several countries are performing pooled sample testing for COVID-19, however, none of the available RT-PCR kits has been tested for pooled sample either by kit manufacturer or research groups. Thus, this study was planned to compare and evaluate seven commercially available COVID-19 RT-PCR kits for pooled samples' testing (Table 1) . Before conducting the study, a survey was done on commercially available RT-PCR kits regarding necessary approvals (US-FDA/CE/ ICMR, India), the lower limit of detection, usage, and availability in India, compatibility with different PCR platforms, cost, and so forth. Based on the survey results, the following kits were selected and Positive pools were created using 60 μl VTM from an RT-PCR con- Amidst the ongoing COVID-19 pandemic, the World Health Organization has globally emphasized the importance of the molecular diagnosis of SARS CoV-2 to limit the spread as well as to appropriately treat those patients who have a serious infection. 12 We conclude that all seven commercially available RT-PCR kits included in this study can be used for the molecular diagnosis of COVID-19. When performing pool sample testing, it might be advisable to use those kits that performed best regarding positive identification in the samples pool that is the TRUPCR SARS-CoV-2 Kit, TaqPath RT-PCR COVID-19 Kit, Allplex 2019-nCOV Assay, and Real-time Fluorescent RT-PCR kit for detecting SARS CoV-2 (BGI). We would like to thank Mr. VK Mishra and Mr. Hemant Verma for their technical help in molecular work. 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Interim guidance 6 Evaluation of COVID-19 RT-qPCR test in multi-sample pools Reg guidance on pooling samples for testing 001 Comparison of commercial RT-PCR diagnostic kits for COVID-19 Comparison of four molecular in vitro diagnostic assays for the detection of SARS-CoV-2 in nasopharyngeal specimens Pooling of SARS-CoV-2 samples to increase molecular testing throughput Evaluation of seven commercial RT-PCR kits for COVID-19 testing in pooled clinical specimens The authors declare that there are no conflict of interests. Atul Garg designed the study, Ujjala Ghoshal prepared and edited the manuscript, Sangram S. Patel, D.V. Singh, and Akshay K. Arya performed the molecular lab work and Shruthi Vasanth performed data analysis. The data that support the findings of this study are available from the corresponding author upon reasonable request. https://orcid.org/0000-0002-7736-5644