Use of RT-PCR thermocycling program for the quantification of DNA viruses in a single run with RNA viruses: Example of Altona RealStar® HSV or VZV PCR kits - 20/11/22
Highlights |
• | Molecular biology is used to detect and quantify DNA and RNA viruses. |
• | PCR thermocycling programs differ for the detection and quantification of DNA and RNA. |
• | The RNA thermocycling program was efficient to quantify DNA of VZV and HSVs. |
• | This method offers the possibility to quantify both DNA and RNA viruses. |
• | This should save technical time and improve laboratory management. |
Abstract |
Real-time PCR plays a key role in the diagnosis of viral infections. Multiple kits can detect or quantify genomes of various viruses with the same thermocycling program. Detection of RNA viruses includes an additional step of reverse transcription and challenge their detection in a single run with DNA viruses.
We investigated the analytical performance of HSV-1, HSV-2 and VZV DNA quantification with Altona RealStar® PCR kits using the RT-PCR program for RNA viruses instead of the PCR program for DNA viruses. For each three viruses, Bland-Altman distribution did not show differences between both programs, and quantification curves generated with both thermocycling programs confirmed high correlation (R2 ≥ 0.9983). Detection of low viral load samples was evaluated, on 10-times repeat-test. All replicate samples were detected with both thermocycling programs and were quantified at similar viral loads (bias in log10 copies/mL: +0.05 (HSV-1), −0.01 (HSV-2) and +0.25 (VZV)). This confirms the feasibility of using the RT-PCR thermocycling program to detect and quantify the genome of RNA and DNA viruses in a single run.
Le texte complet de cet article est disponible en PDF.Keywords : Real-Time PCR, Thermocycling program, RNA viruses, DNA viruses
Plan
Vol 52 - N° 8
P. 453-455 - novembre 2022 Retour au numéro
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