Key result
Quantitative Taqman PCR assay detects HIV-1 proviral DNA down to 10 copies per reaction.
Why the study?
Quantification of HIV-1 proviral DNA in newly infected individuals is important for monitoring infection but requires sensitive and reliable assays.
Does the Taqman Real-Time PCR Assay accurately quantify HIV Proviral DNA in newly infected HIV-1 individuals?
Does the Taqman Real-Time PCR Assay accurately quantify HIV Proviral DNA in newly infected HIV-1 individuals?
The Taqman Real-Time PCR Assay demonstrates good analytic sensitivity and a wide linear dynamic range for quantifying HIV proviral DNA, supporting its use in routine diagnostics.
Offers a sensitive tool for HIV-1 proviral DNA quantification; supports further research but should not yet alter clinical practice.
Human Immunodeficiency Virus (HIV) is a virus belonging to the family Retroviridae. HIV – 1 is found to be predominant in India and many parts of Africa. The intention of this study was to quantify the HIV Proviral Deoxyribonucleic Acid (DNA) from newly infected HIV-1 individuals. Fifty patients who were tested positive for HIV were included in this study. Proviral Ribo Nucleic Acid (RNA) was extracted by QIAmp® RNA Mini Kit (QIAGEN, Germany) method. Complementary Deoxyribo Nucleic Acid (cDNA) was synthesized by using Invitrogen Superscript III cDNA synthesis Kit (USA). This cDNA was subjected to Polymerase Chain Reaction (PCR) and Gene cloning by transformation method. The quantification of Real time PCR was done by Applied Bio-System (ABI)-Prism 7700. A linear standard curve was obtained 10 copies to 106 copies per reaction. The assay had good analytic sensitivity and linear dynamic range greater than 6 logs. From the results obtained in this study, It was concluded that Taqman Real-Time PCR Assay plays a major role in monitoring the HIV infected patients in routine diagnostics and clinical practice.
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Elumalai et al. (2022) studied HIV-1 infection (n=50). Quantitative Taqman Real-Time PCR Assay was evaluated on Analytic sensitivity and linear dynamic range. The quantitative Taqman Real-Time PCR assay demonstrated a detection limit of 10 copies per reaction and a linear dynamic range greater than 6 logs for measuring HIV-1 proviral DNA.
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