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Pooled RNA sample reverse transcriptase real time PCR assay for SARS CoV-2 infection: A reliable, faster and economical method.
PLoS One. 2020; 15(7):e0236859.Plos

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+Show Affiliations

Department of Clinical Virology, Institute of Liver and Biliary Sciences, New Delhi, India.Department of Clinical Virology, Institute of Liver and Biliary Sciences, New Delhi, India.Department of Clinical Virology, Institute of Liver and Biliary Sciences, New Delhi, India.Department of Clinical Virology, Institute of Liver and Biliary Sciences, New Delhi, India.Department of Clinical Virology, Institute of Liver and Biliary Sciences, New Delhi, India.Department of Clinical Virology, Institute of Liver and Biliary Sciences, New Delhi, India.Department of Clinical Virology, Institute of Liver and Biliary Sciences, New Delhi, India.Department of Hospital Operations, Institute of Liver and Biliary Sciences, New Delhi, India.Department of Clinical Research, Institute of Liver and Biliary Sciences, New Delhi, India.Department of Hepatology, Institute of Liver and Biliary Sciences, New Delhi, India.

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

32730368

Citation

Gupta, Ekta, et al. "Pooled RNA Sample Reverse Transcriptase Real Time PCR Assay for SARS CoV-2 Infection: a Reliable, Faster and Economical Method." PloS One, vol. 15, no. 7, 2020, pp. e0236859.
Gupta E, Padhi A, Khodare A, et al. Pooled RNA sample reverse transcriptase real time PCR assay for SARS CoV-2 infection: A reliable, faster and economical method. PLoS One. 2020;15(7):e0236859.
Gupta, E., Padhi, A., Khodare, A., Agarwal, R., Ramachandran, K., Mehta, V., Kilikdar, M., Dubey, S., Kumar, G., & Sarin, S. K. (2020). Pooled RNA sample reverse transcriptase real time PCR assay for SARS CoV-2 infection: A reliable, faster and economical method. PloS One, 15(7), e0236859. https://doi.org/10.1371/journal.pone.0236859
Gupta E, et al. Pooled RNA Sample Reverse Transcriptase Real Time PCR Assay for SARS CoV-2 Infection: a Reliable, Faster and Economical Method. PLoS One. 2020;15(7):e0236859. PubMed PMID: 32730368.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Pooled RNA sample reverse transcriptase real time PCR assay for SARS CoV-2 infection: A reliable, faster and economical method. AU - Gupta,Ekta, AU - Padhi,Abhishek, AU - Khodare,Arvind, AU - Agarwal,Reshu, AU - Ramachandran,Krithiga, AU - Mehta,Vibha, AU - Kilikdar,Mousumi, AU - Dubey,Shantanu, AU - Kumar,Guresh, AU - Sarin,Shiv Kumar, Y1 - 2020/07/30/ PY - 2020/05/05/received PY - 2020/07/15/accepted PY - 2020/7/31/entrez PY - 2020/7/31/pubmed PY - 2020/8/22/medline SP - e0236859 EP - e0236859 JF - PloS one JO - PLoS One VL - 15 IS - 7 N2 - 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. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/32730368/Pooled_RNA_sample_reverse_transcriptase_real_time_PCR_assay_for_SARS_CoV_2_infection:_A_reliable_faster_and_economical_method_ L2 - https://dx.plos.org/10.1371/journal.pone.0236859 DB - PRIME DP - Unbound Medicine ER -