Establishment of cheap and reliable real-time PCR for quantitation of HIV-1 viral load in plasma

Objective: To establish an inexpensive and reliable real-time PCR for quantitation of HIV-1 RNA from plasma samples. Material and Method: Previously analyzed 145 HIV-1 positive plasma samples with viral load ranging from less than 40 to approximately 1,000,000 copies/ml were included in the present...

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Main Authors: Supadej,K., Intorasoot,S.
Format: Article
Published: Medical Association of Thailand 2015
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Online Access:http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84871692946&origin=inward
http://cmuir.cmu.ac.th/handle/6653943832/38109
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-381092015-06-16T07:38:25Z Establishment of cheap and reliable real-time PCR for quantitation of HIV-1 viral load in plasma Supadej,K. Intorasoot,S. Medicine (all) Objective: To establish an inexpensive and reliable real-time PCR for quantitation of HIV-1 RNA from plasma samples. Material and Method: Previously analyzed 145 HIV-1 positive plasma samples with viral load ranging from less than 40 to approximately 1,000,000 copies/ml were included in the present study. HIV-1 gag gene was amplified and cloned into TA cloning vector. External standard curve was plotted using in vitro transcribed HIV-1 RNA and utilized for viral quantitation in the samples. Scramble nucleotides located in HIV-1 specific probe was subsequently constructed and used for individual systemic control. The correlation coefficient and Bland-Altman plot were applied for statistical analysis of the two methods. Results: The limit of quantitation of the validated assay was 31 copies/ml and the linear range was approximate 31-1 × 107 copies/ml. After reproducibility determination using intra-and inter-run assay, it was implied that the coefficient of variation (%CV) was significantly increased while the low copy number of RNA was examined. A highly correlation (r2 = 0.8099) and good agreement were obtained when the two assays were compared. Conclusion: Developed real-time PCR was inexpensive and reliable for quantitation of HIV-1 viral load in plasma. 2015-06-16T07:38:25Z 2015-06-16T07:38:25Z 2012-12-01 Article 01252208 2-s2.0-84871692946 23390788 http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84871692946&origin=inward http://cmuir.cmu.ac.th/handle/6653943832/38109 Medical Association of Thailand
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Medicine (all)
spellingShingle Medicine (all)
Supadej,K.
Intorasoot,S.
Establishment of cheap and reliable real-time PCR for quantitation of HIV-1 viral load in plasma
description Objective: To establish an inexpensive and reliable real-time PCR for quantitation of HIV-1 RNA from plasma samples. Material and Method: Previously analyzed 145 HIV-1 positive plasma samples with viral load ranging from less than 40 to approximately 1,000,000 copies/ml were included in the present study. HIV-1 gag gene was amplified and cloned into TA cloning vector. External standard curve was plotted using in vitro transcribed HIV-1 RNA and utilized for viral quantitation in the samples. Scramble nucleotides located in HIV-1 specific probe was subsequently constructed and used for individual systemic control. The correlation coefficient and Bland-Altman plot were applied for statistical analysis of the two methods. Results: The limit of quantitation of the validated assay was 31 copies/ml and the linear range was approximate 31-1 × 107 copies/ml. After reproducibility determination using intra-and inter-run assay, it was implied that the coefficient of variation (%CV) was significantly increased while the low copy number of RNA was examined. A highly correlation (r2 = 0.8099) and good agreement were obtained when the two assays were compared. Conclusion: Developed real-time PCR was inexpensive and reliable for quantitation of HIV-1 viral load in plasma.
format Article
author Supadej,K.
Intorasoot,S.
author_facet Supadej,K.
Intorasoot,S.
author_sort Supadej,K.
title Establishment of cheap and reliable real-time PCR for quantitation of HIV-1 viral load in plasma
title_short Establishment of cheap and reliable real-time PCR for quantitation of HIV-1 viral load in plasma
title_full Establishment of cheap and reliable real-time PCR for quantitation of HIV-1 viral load in plasma
title_fullStr Establishment of cheap and reliable real-time PCR for quantitation of HIV-1 viral load in plasma
title_full_unstemmed Establishment of cheap and reliable real-time PCR for quantitation of HIV-1 viral load in plasma
title_sort establishment of cheap and reliable real-time pcr for quantitation of hiv-1 viral load in plasma
publisher Medical Association of Thailand
publishDate 2015
url http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84871692946&origin=inward
http://cmuir.cmu.ac.th/handle/6653943832/38109
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