Analytical performance and standardization of four HCV RNA assays in China: an evaluation of sensitivity, precision, and genotype inclusivity

IntroductionAccurate, specific, and sensitive detection and quantification of hepatitis C virus (HCV) RNA are critical for diagnosing and managing HCV infections. This study evaluated and compared the performance of four commercially available HCV RNA quantification reagents using standardized serum...

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Main Authors: Hao Yan, Bing Xu, Qinyi Zhang, Lijun Zhang, Zhihan Fang, Huijun Zhang, Junhang Pan, Wenge Xing
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-06-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1594410/full
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author Hao Yan
Hao Yan
Bing Xu
Qinyi Zhang
Qinyi Zhang
Lijun Zhang
Zhihan Fang
Huijun Zhang
Junhang Pan
Wenge Xing
author_facet Hao Yan
Hao Yan
Bing Xu
Qinyi Zhang
Qinyi Zhang
Lijun Zhang
Zhihan Fang
Huijun Zhang
Junhang Pan
Wenge Xing
author_sort Hao Yan
collection DOAJ
description IntroductionAccurate, specific, and sensitive detection and quantification of hepatitis C virus (HCV) RNA are critical for diagnosing and managing HCV infections. This study evaluated and compared the performance of four commercially available HCV RNA quantification reagents using standardized serum panels, providing evidence-based insights for clinical applications.MethodsPerformance metrics, including analytical sensitivity, specificity, limit of detection (LOD), precision, genotype inclusivity (GT 1–6), and linearity, were assessed using seven distinct serum panels: basic, analytical specificity, seroconversion, analytical sensitivity, precision, genotype qualification, and linearity panels.ResultsAll reagents demonstrated 100% analytical sensitivity and specificity (95% CI: 79.95–100), with no cross-reactivity to common interfering substances or viruses. LOD values for reagents A, B, C, and D were 25, 50, 50, and 50 IU/ml, respectively. Intra- and inter-assay coefficients of variation (CVs) for HCV genotypes 1–6 ranged from 1.48 to 4.37% and 1.74 to 4.84%, respectively. Strong linear correlations (R2 > 0.95) were observed between measured and expected HCV RNA levels across all reagents.ConclusionThese reagents exhibit high sensitivity, specificity, precision, and accuracy for HCV genotypes 1–6, with a wide linear range, making them suitable for clinical diagnosis and monitoring of HCV infections.
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spelling doaj-art-9d4617063e894723979d3a97542817f22025-08-20T03:18:44ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-06-011610.3389/fmicb.2025.15944101594410Analytical performance and standardization of four HCV RNA assays in China: an evaluation of sensitivity, precision, and genotype inclusivityHao Yan0Hao Yan1Bing Xu2Qinyi Zhang3Qinyi Zhang4Lijun Zhang5Zhihan Fang6Huijun Zhang7Junhang Pan8Wenge Xing9Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, ChinaKey Laboratory of Public Health Detection and Etiological Research of Zhejiang Province, Hangzhou, ChinaNational HIV/AIDS Reference Laboratory, National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, ChinaZhejiang Provincial Center for Disease Control and Prevention, Hangzhou, ChinaFuyang District Center for Disease Control and Prevention, Hangzhou, ChinaMinhang Hospital, Fudan University, Shanghai, ChinaYangpu Hospital, Tongji University, Shanghai, ChinaInstitute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, National Human Diseases Animal Model Resource Center, Beijing, ChinaZhejiang Provincial Center for Disease Control and Prevention, Hangzhou, ChinaNational HIV/AIDS Reference Laboratory, National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, ChinaIntroductionAccurate, specific, and sensitive detection and quantification of hepatitis C virus (HCV) RNA are critical for diagnosing and managing HCV infections. This study evaluated and compared the performance of four commercially available HCV RNA quantification reagents using standardized serum panels, providing evidence-based insights for clinical applications.MethodsPerformance metrics, including analytical sensitivity, specificity, limit of detection (LOD), precision, genotype inclusivity (GT 1–6), and linearity, were assessed using seven distinct serum panels: basic, analytical specificity, seroconversion, analytical sensitivity, precision, genotype qualification, and linearity panels.ResultsAll reagents demonstrated 100% analytical sensitivity and specificity (95% CI: 79.95–100), with no cross-reactivity to common interfering substances or viruses. LOD values for reagents A, B, C, and D were 25, 50, 50, and 50 IU/ml, respectively. Intra- and inter-assay coefficients of variation (CVs) for HCV genotypes 1–6 ranged from 1.48 to 4.37% and 1.74 to 4.84%, respectively. Strong linear correlations (R2 > 0.95) were observed between measured and expected HCV RNA levels across all reagents.ConclusionThese reagents exhibit high sensitivity, specificity, precision, and accuracy for HCV genotypes 1–6, with a wide linear range, making them suitable for clinical diagnosis and monitoring of HCV infections.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1594410/fullhepatitis C virusRNAperformance assessmentquantitativeserum panel
spellingShingle Hao Yan
Hao Yan
Bing Xu
Qinyi Zhang
Qinyi Zhang
Lijun Zhang
Zhihan Fang
Huijun Zhang
Junhang Pan
Wenge Xing
Analytical performance and standardization of four HCV RNA assays in China: an evaluation of sensitivity, precision, and genotype inclusivity
Frontiers in Microbiology
hepatitis C virus
RNA
performance assessment
quantitative
serum panel
title Analytical performance and standardization of four HCV RNA assays in China: an evaluation of sensitivity, precision, and genotype inclusivity
title_full Analytical performance and standardization of four HCV RNA assays in China: an evaluation of sensitivity, precision, and genotype inclusivity
title_fullStr Analytical performance and standardization of four HCV RNA assays in China: an evaluation of sensitivity, precision, and genotype inclusivity
title_full_unstemmed Analytical performance and standardization of four HCV RNA assays in China: an evaluation of sensitivity, precision, and genotype inclusivity
title_short Analytical performance and standardization of four HCV RNA assays in China: an evaluation of sensitivity, precision, and genotype inclusivity
title_sort analytical performance and standardization of four hcv rna assays in china an evaluation of sensitivity precision and genotype inclusivity
topic hepatitis C virus
RNA
performance assessment
quantitative
serum panel
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1594410/full
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