SARS-CoV-2 Direct Real Time PCR without RNA Extraction: Appropriate Method or Not?

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  •   Spandan Chaudhary

  •   Disha Patel

  •   Kavisha Vyas

  •   Pooja Chaudhary

  •   Prathana Singhania

  •   Juhi Patel

  •   Ekta Jajodia

  •   Nilay Dave

  •   Shiv Patel

  •   Ramachandiran Sivaramakrishnan

  •   Mayank Patadiya

  •   Tushar Sonagara

  •   Ashish Hirpara

  •   Neeraj Arora

Abstract


Real time PCR (RT-PCR) detection method is the widely used for COVID-19 virus detection. This includes sample collection in viral transport medium (VTM), viral RNA extraction followed by detection of virus using fluorescence dye-based system using RT-PCR machine. Several studies have demonstrated a new method which replaces the extraction step by a simple method involving DTT and Proteinase-K and heat treatment. ICMR and few other governing bodies have approved such protocols but are they appropriate in clinical context? In present study, we tried to evaluate one such protocol by using ICMR and WHO approved COVID-19 detection protocol (of CoviPath™ COVID-19 RT-PCR Kit) by replacing RNA extraction step. We used 228 clinical COVID-19 samples for studying method which includes 176 positive (CT values from 14 to 23; 24 to 31 and 32 to 37 were considered as high, moderate and low positive respectively) and 52 negative nasopharyngeal and oropharyngeal swabs samples. We got 100% concordant results with negative samples and 92% concordant and 8% non-concordant results for positive samples. Non-concordant results are with low positive samples. Low level of positivity in the samples could indicate the initial/end stage of COVID-19 disease. If they are at the initial stage, they can be the potential carrier and spread the disease. Authors believe that direct methods can be used for screening bit not for diagnosis of COVID-19 disease.



Keywords: COVID-19, Direct RT-PCR, DTT, Proteinase-K

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How to Cite
Chaudhary, S., Patel, D., Vyas, K., Chaudhary, P., Singhania, P., Patel, J., Jajodia, E., Dave, N., Patel, S., Sivaramakrishnan, R., Patadiya, M., Sonagara, T., Hirpara, A., & Arora, N. (2022). SARS-CoV-2 Direct Real Time PCR without RNA Extraction: Appropriate Method or Not?. European Journal of Medical and Health Sciences, 4(1), 41–47. https://doi.org/10.24018/ejmed.2022.4.1.1164