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Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum

BACKGROUND: The duplex mutation primers offer many advantages over other multi-labeled probes for real-time detection of amplification products. OBJECTIVES: To develop and validate a novel real-time PCR for quantification of HCV RNA based on the duplex mutation primers technology. MATERIALS AND METH...

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Autores principales: Xia, Qian-Feng, Wen, Yang-An, Liu, Ping, Li, Pu, Liu, Jin-Bo, Qin, Xi, Qian, Shi-Yun, Tu, Zhi-Guang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Kowsar 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3212762/
https://www.ncbi.nlm.nih.gov/pubmed/22087189
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author Xia, Qian-Feng
Wen, Yang-An
Liu, Ping
Li, Pu
Liu, Jin-Bo
Qin, Xi
Qian, Shi-Yun
Tu, Zhi-Guang
author_facet Xia, Qian-Feng
Wen, Yang-An
Liu, Ping
Li, Pu
Liu, Jin-Bo
Qin, Xi
Qian, Shi-Yun
Tu, Zhi-Guang
author_sort Xia, Qian-Feng
collection PubMed
description BACKGROUND: The duplex mutation primers offer many advantages over other multi-labeled probes for real-time detection of amplification products. OBJECTIVES: To develop and validate a novel real-time PCR for quantification of HCV RNA based on the duplex mutation primers technology. MATERIALS AND METHODS: The duplex mutation primers were selected in the highly conservative 5' non-coding region (5'NCR) of the HCV RNA. The assay was validated with the Viral Quality Control panel, which also includes Chinese HCV RNA standards. RESULTS: The detection limit was 57 IU/mL, and a good linear correlation in the range of 102-108 IU/mL was revealed (r(2) = 0.999) with the novel method. This assay has a dynamic range of at least 8 log10 without the need for specimen dilution, good clinical intra- and inter-run precision, and excellent correlation with a commercially available assay(r(2) = 0.95). CONCLUSIONS: The high sensitivity, wide linear range, and good reproducibility, combined with low cost, make this novel quantitative HCV real-time PCR assay particularly well suited for application to clinical and epidemiological studies.
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spelling pubmed-32127622011-11-15 Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum Xia, Qian-Feng Wen, Yang-An Liu, Ping Li, Pu Liu, Jin-Bo Qin, Xi Qian, Shi-Yun Tu, Zhi-Guang Hepat Mon Original Article BACKGROUND: The duplex mutation primers offer many advantages over other multi-labeled probes for real-time detection of amplification products. OBJECTIVES: To develop and validate a novel real-time PCR for quantification of HCV RNA based on the duplex mutation primers technology. MATERIALS AND METHODS: The duplex mutation primers were selected in the highly conservative 5' non-coding region (5'NCR) of the HCV RNA. The assay was validated with the Viral Quality Control panel, which also includes Chinese HCV RNA standards. RESULTS: The detection limit was 57 IU/mL, and a good linear correlation in the range of 102-108 IU/mL was revealed (r(2) = 0.999) with the novel method. This assay has a dynamic range of at least 8 log10 without the need for specimen dilution, good clinical intra- and inter-run precision, and excellent correlation with a commercially available assay(r(2) = 0.95). CONCLUSIONS: The high sensitivity, wide linear range, and good reproducibility, combined with low cost, make this novel quantitative HCV real-time PCR assay particularly well suited for application to clinical and epidemiological studies. Kowsar 2011-07-01 2011-07-01 /pmc/articles/PMC3212762/ /pubmed/22087189 Text en Copyright © 2011, Kowsar M.P. Co. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Xia, Qian-Feng
Wen, Yang-An
Liu, Ping
Li, Pu
Liu, Jin-Bo
Qin, Xi
Qian, Shi-Yun
Tu, Zhi-Guang
Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum
title Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum
title_full Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum
title_fullStr Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum
title_full_unstemmed Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum
title_short Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum
title_sort use of duplex mutation primers for real-time pcr quantification of hepatitis c virus rna in serum
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3212762/
https://www.ncbi.nlm.nih.gov/pubmed/22087189
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