• Ankesh Gupta, Ankit Mittal, Shivram Dhakad, Megha Brijwal, Manish Soneja, Deepankar Srigyan, Ashit Xess, Arvind Kumar, Animesh Ray, Ved Prakash Meena, Rohit Kumar Garg, Komal Singh, Devashish Desai, Mohit Chowdhury, Souradeep Chowdhury, Kunal Sharma, Ananthu Narayan, G Rahul Krishnan, Shivdas Naik, Lalit Dar, Ravindra Mohan Pandey, Shivam Pandey, Sanjeev Sinha, and Naveet Wig.
• Department of Medicine, All India Institute of Medical Sciences, New Delhi, India.
• Indian J Med Res. 2021 May 1; 153 (5&6): 665-670.

Background & ObjectivesIn the present scenario, the most common sample for diagnosis of COVID-19 by reverse transcription polymerase chain reaction (RT-PCR) is nasal and throat swab (NTS). Other sampling options such as gargle lavage have found limited application in clinical use mostly because of unavailability of an appropriate gargling liquid. This study was conducted to assess the stability of SARS-CoV-2 RNA in normal saline at 4°C that can serve as a gargling liquid as well as a transport medium. The study also looked at the agreement between NTS and gargle lavage/saliva for the detection of SARS-CoV-2.MethodsIn 29 consecutive real-time RT-PCR (rRT-PCR) positive COVID-19 patients, paired NTS, gargle and saliva samples were taken. Samples were processed by rRT-PCR for the detection of SARS-CoV-2 RNA. To assess the SARS-CoV-2 RNA stability in normal saline, gargle lavage specimens were divided into two aliquots; one subset of the specimen was run within 4-6 h along with the routine samples (NTS and saliva) and the other subset was stored at 4°C and processed after 24-30 h. Agreement between cycle threshold (Ct) values from both the runs was compared using Bland-Altman (BA) analysis.ResultsThe positivity rates of rRT-PCR in NTS, saliva and gargle lavage samples were 82.7 (24/29), 79.3 (23/29) and 86.2 per cent (25/29), respectively. BA plot showed a good agreement between the Ct values of fresh and stored gargle samples, stipulating that there were no significant differences in the approximate viral load levels between the fresh and stored gargle lavage samples (bias: E gene -0.64, N gene -0.51, ORF gene -0.19).Interpretation & ConclusionsOur study results show stability of SARS-CoV-2 RNA in the gargle samples collected using normal saline up to 24-30 h. Gargle lavage and saliva specimen collection are cost-effective and acceptable methods of sampling for the detection of SARS-CoV-2 RNA by rRT-PCR. These simplified, inexpensive and acceptable methods of specimen collection would reduce the cost and workload on healthcare workers for sample collection.

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