Evaluation of detection of severe acute respiratory syndrome coronavirus-2 by chip-based real-time polymerase chain reaction test (truenat™ beta CoV) in multi-sample pools
Venkata Giri Prasad Polu1, Neela Mani kanta Kota2, Deepthi Karumanchi3, Sreekanth Reddy Basireddy4, Sandhya Munagapati5, Shiva Kumar Mugudalabetta1, Venkata Prasad Ganta6, Uday Sankar Allam7
1 Damien Foundation India Trust, Vikrama Simhapuri University, Nellore, Andhra Pradesh, India
2 Department of Tourism Management, Vikrama Simhapuri University, Nellore, Andhra Pradesh, India
3 Department of Microbiology, Krishna Institute of Medical Sciences (KIMS), Nellore, Andhra Pradesh, India
4 Department of Microbiology, A.C. Subbareddy Govt. Medical College, Nellore, Andhra Pradesh, India
5 Scientist, Crop Production, DATTC Center, ANGRAU, Nellore, Andhra Pradesh, India
6 District TB Officer, Vikrama Simhapuri University, Nellore, Andhra Pradesh, India
7 Department of Biotechnology, Vikrama Simhapuri University, Nellore, Andhra Pradesh, India
Dr. Uday Sankar Allam
Department of Biotechnology, Vikrama Simhapuri University, Nellore - 524 324, Andhra Pradesh
Source of Support: None, Conflict of Interest: None
Introduction: Systematic testing for Severe Acute Respiratory Syndrome CoronaVirus-2 (SARS-CoV-2) using molecular diagnostic tools to identify individuals with coronavirus disease 2019 (COVID-19) infection, and tracing their primary and secondary contacts is important to curb its spread. With resource limitations on testing individual samples, testing of pooled samples provides alternative approach to increase testing capacity. Present aimed at assessing the detection of SARS-CoV-2 RNA in pooled samples using chip-based real-time polymerase chain reaction Test (Truenat™ Beta CoV).
Materials and Methods: Pooled sample size of five was used from laboratory confirmed COVID-19 positive and negative samples. SARS-CoV-2 positive nasopharyngeal specimens of known samples from high, medium, low, and very low viral load were mixed with SARS-CoV-2 negative nasopharyngeal specimens of known samples in 1:4 ratio, followed by analysis using Truenat. Furthermore, each sample in that pool was tested individually. Pooled sample testing was also done on the samples of unknown status.
Results: The results of the present study showed cycle threshold (Ct) values of pooled sample with SARS-CoV-2 positive RNA of high, medium, low, and very low viral load were 16.8, 24.22, 28.2, and 33.43, compared to Ct values of individual samples of 16.43, 22.0, 28.00, and 33.00, respectively.
Conclusion: These results suggest that the Ct values of pooled samples were in agreement with Ct values of individual samples indicating the validity of pooled sample testing for screening SARS-CoV-2 using Truenat.
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