Pooled RNA sample reverse transcriptase real time PCR assay for SARS CoV-2 infection: A reliable, faster and economical method.

Abstract

BACKGROUND:

Corona virus disease 2019 (COVID-19) which initially started as a cluster of pneumonia cases in the Wuhan city of China has now become a full-blown pandemic. Timely diagnosis of COVID-19 is the key in containing the pandemic and breaking the chain of transmission. In low- and middle-income countries availability of testing kits has become the major bottleneck in testing. Novel methods like pooling of samples are the need of the hour.

OBJECTIVE:

We undertook this study to evaluate a novel protocol of pooling of RNA samples/elutes in performance of PCR for SARS CoV-2 virus.

STUDY DESIGN:

Extracted RNA samples were randomly placed in pools of 8 on a 96 well plate. Both individual RNA (ID) and pooled RNA RT-qPCR for the screening E gene were done in the same plate and the positivity for the E gene was seen.

RESULTS:

The present study demonstrated that pool testing with RNA samples can easily detect even up to a single positive sample with Ct value as high as 38. The present study also showed that the results of pool testing is not affected by number of positive samples in a pool.

CONCLUSION:

Pooling of RNA samples can reduce the time and expense, and can help expand diagnostic capabilities, especially during constrained supply of reagents and PCR kits for the diagnosis of SARS-CoV-2 infection.

• Authors: Ekta Gupta, Abhishek Padhi, Arvind Khodare, Reshu Agarwal, Krithiga Ramachandran, Vibha Mehta, Mousumi Kilikdar, Shantanu Dubey, Guresh Kumar, Shiv Kumar Sarin
• Journal: PloS one (PLoS One) Vol. 15 Issue 7 Pg. e0236859 ( 2020) ISSN: 1932-6203 [Electronic] United States
• PMID: 32730368 (Publication Type: Evaluation Study, Journal Article)

Chemical References

• Coronavirus Envelope Proteins
• RNA, Viral
• Viral Envelope Proteins
• envelope protein, SARS-CoV-2

Topics

• Betacoronavirus (genetics)
• COVID-19
• Coronavirus Envelope Proteins
• Coronavirus Infections (diagnosis, epidemiology, virology)
• Diagnostic Tests, Routine (economics, methods)
• Humans
• India (epidemiology)
• Mass Screening (economics, methods)
• Pandemics
• Pneumonia, Viral (diagnosis, epidemiology, virology)
• Prospective Studies
• RNA, Viral (genetics, isolation & purification)
• Real-Time Polymerase Chain Reaction (economics, methods)
• Reproducibility of Results
• Reverse Transcriptase Polymerase Chain Reaction (economics, methods)
• SARS-CoV-2
• Viral Envelope Proteins (genetics)