Research Article

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

Spandan Chaudhary
Unipath Specialty Laboratory Ltd, India
* Corresponding author
Disha Patel
Unipath Specialty Laboratory Ltd, India
Kavisha Vyas
Unipath Specialty Laboratory Ltd, India
Pooja Chaudhary
Unipath Specialty Laboratory Ltd, India
Prathana Singhania
Unipath Specialty Laboratory Ltd, India
Juhi Patel
Unipath Specialty Laboratory Ltd, India
Ekta Jajodia
Unipath Specialty Laboratory Ltd, India
Nilay Dave
Unipath Specialty Laboratory Ltd, India
Shiv Patel
Unipath Specialty Laboratory Ltd, India
Ramachandiran Sivaramakrishnan
Unipath Specialty Laboratory Ltd, India
Mayank Patadiya
Unipath Specialty Laboratory Ltd, India
Tushar Sonagara
Unipath Specialty Laboratory Ltd, India
Ashish Hirpara
Unipath Specialty Laboratory Ltd, India
Neeraj Arora
Unipath Specialty Laboratory Ltd, India
DOI icon 10.24018/ejmed.2022.4.1.1164

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Submitted 2021-11-30
Published 2022-01-27

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Table of Contents

Article Main Content

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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