BACKGROUND: Most of the quantitation assays for HIV-1 RNA used currently are designed and optimized for HIV-1 subtype B viruses and hence may not be suitable for India, where the predominant subtype is HIV-1 subtype C. OBJECTIVES: Development and standardization of HIV-1 TaqMan real time PCR assay suitable for measuring plasma and CSF viral RNA levels in HIV subtype C infected individuals. STUDY DESIGN: A TaqMan real time PCR was developed using primers and probes selected in the gag region for detection of Indian HIV-1 subtype C strain. Plasma (n=120) and CSF samples (n=46) obtained from HIV infected subjects were used to evaluate the sensitivity and specificity of the assay. A comparative evaluation was carried out with a commercially available quantitative HIV viral load assay (Roche Amplicor Version 1.5). RESULTS: The TaqMan assay was able to amplify all HIV-1 group M subtypes except subtype E. Viral loads could be estimated in all the plasma (n=120) and 40/46 CSF samples obtained from HIV positive subjects. Sensitivity of this assay was found to be 180 copies/ml. Correlation with the commercially available viral load assay was very good (r=0.885). CONCLUSIONS: A TaqMan real time PCR was standardized for HIV-1 subtype C and it was more sensitive (180 copies/ml) than standard Amplicor monitor assay, Version 1.5 (400 copies/ml).
BACKGROUND: Most of the quantitation assays for HIV-1 RNA used currently are designed and optimized for HIV-1 subtype B viruses and hence may not be suitable for India, where the predominant subtype is HIV-1 subtype C. OBJECTIVES: Development and standardization of HIV-1 TaqMan real time PCR assay suitable for measuring plasma and CSF viral RNA levels in HIV subtype C infected individuals. STUDY DESIGN: A TaqMan real time PCR was developed using primers and probes selected in the gag region for detection of Indian HIV-1 subtype C strain. Plasma (n=120) and CSF samples (n=46) obtained from HIV infected subjects were used to evaluate the sensitivity and specificity of the assay. A comparative evaluation was carried out with a commercially available quantitative HIV viral load assay (Roche Amplicor Version 1.5). RESULTS: The TaqMan assay was able to amplify all HIV-1 group M subtypes except subtype E. Viral loads could be estimated in all the plasma (n=120) and 40/46 CSF samples obtained from HIV positive subjects. Sensitivity of this assay was found to be 180 copies/ml. Correlation with the commercially available viral load assay was very good (r=0.885). CONCLUSIONS: A TaqMan real time PCR was standardized for HIV-1 subtype C and it was more sensitive (180 copies/ml) than standard Amplicor monitor assay, Version 1.5 (400 copies/ml).
Authors: Peilin Li; Theodore Ruel; Katsuya Fujimoto; Hiroyu Hatano; Steven Yukl; Leigh Anne Eller; Teri Liegler; Moses Kamya; Anne Gassasira; Grant Dorsey; Philip J Rosenthal; Diane V Havlir; Joseph K Wong Journal: J Virol Methods Date: 2010-09-21 Impact factor: 2.014
Authors: Cecile A Delille; Sarah T Pruett; Vincent C Marconi; Jeffrey L Lennox; Wendy S Armstrong; Richard F Arrendale; Anandi N Sheth; Kirk A Easley; Edward P Acosta; Aswani Vunnava; Ighovwerha Ofotokun Journal: J Clin Pharmacol Date: 2014-04-08 Impact factor: 3.126
Authors: K Gopukumar; Shobini L Rao; P Satishchandra; Jayashree Dasgupta; Ronald J Ellis; D K Subbakrishna; P Mariamma; Anupa Kamat; Anita Desai; V Ravi; B S Rao; K S Satish; Mahendra Kumar Journal: J Neurovirol Date: 2008-11 Impact factor: 2.643