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New RT-PCR kit for accurate detection of various mutant strains

Various studies have shown that RdRp and ORF1b-nsp14 genes are more sensitive in detecting Covid-19…reports Asian Lite News

An Institute of National Importance under the Ministry of Science and Technology has developed a new RT-PCR kit with novel gene which targets facilitating detection of Covid-19 across various mutant strains.

In a statement, the Ministry of Science and Technology said, “A newly developed multiplex RT-PCR kit has a higher accuracy of detecting Covid 19 across the various mutant strains of the virus responsible for the global pandemic. As the pandemic is going through a second wave with multiple variants, the selection of target genes in multiplex RT-PCR assay is becoming critical for accurate detection of the virus.”

Even though coronaviruses make far fewer errors than other RNA viruses, the mutations in S, R, and N genes often interfere with RT-PCR assay.

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“The new multiplex RT-PCR kit developed by Sree Chitra Tirunal Institute for Medical Sciences and Technology (SCTIMST), an Institute of National Importance under the Department of Science and Technology, Government of India targets two SARS-CoV-2 genes: RdRp and ORFb-nsp14, and the human RNAse P gene as the internal control to help detect a range of mutant strains,” the Ministry said.

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Various studies have shown that RdRp and ORF1b-nsp14 genes are more sensitive in detecting Covid-19. In order to target the multiple variants in the second wave, using two highly accurate confirmatory genes like RdRp and ORF-nsp14, can give precise results. The ORFb-nsp14 is one of the least mutated genes in Covid-19 and currently there are no kits in the market with ORF-nsp14 as the target.

“The new kit is based on multiplex Taqman chemistry, amplifying all three genes in a single reaction. The amplification time for the assay is 45 minutes, apart from the time required for the RNA isolation from nasopharyngeal swab samples. Multiplexing two confirmatory genes will help shortlist possible new variants if one of the genes fails to amplify and can be marked for sequence analysis,” the Ministry said.

The Indian Council of Medical Research (ICMR) has validated this kit at the National Institute of Virology (NIV) Pune, and found that it has 97.3 per cent sensitivity and 100 per cent specificity in Covid 19 detection.

The SCTIMST has signed a non-exclusive license MoU with Huwel Lifesciences, Hyderabad, on May 14 to commercialize the kit.

“This unique RT-PCR kit will be a significant weapon in our fight against COVID-19 by a facile detection of SARS-CoV-2 mutations which are becoming increasingly important,” said Prof Ashutosh Sharma, Secretary, DST.

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