ISSN: 0973-7510

E-ISSN: 2581-690X

Research Article | Open Access
Suguna Elumalai1, Chitralekha Saikumar2, Florida Tilton3 and Lakshmi Krishnasamy2
1Genomic Research Centre, Sree Balaji Medical College & Hospital, BIHER University, Chromepet, Chennai – 600 044, Tamilnadu, India.
2Department of Microbiology, Sree Balaji Medical College & Hospital, BIHER University, Chromepet, Chennai- 600 044, Tamilnadu, India.
3Molecular Diagnostics, Sree Balaji Medical College & Hospital, BIHER University, Chromepet, Chennai – 600 044, Tamilnadu, India.
J Pure Appl Microbiol. 2022;16(2):1096-1102 | Article Number: 7166
https://doi.org/10.22207/JPAM.16.2.34 | © The Author(s). 2022
Received: 08/07/2021 | Accepted: 28/03/2022 | Published online: 27/05/2022
Issue online: June 2022
Abstract

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.

Keywords

Viral load, HIV-1, Reverse transcriptase, Taqman probe assay, pGEMT-Vector ligation, Gene transformation

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