Elbeik Tarek, Surtihadi Johan, Destree Mark, Gorlin Jed, Holodniy Mark, Jortani Saeed A, Kuramoto Ken, Ng Valerie, Valdes Roland, Valsamakis Alexandra, Terrault Norah A
Department of Laboratory Medicine, University of California, San Francisco 94110, USA.
J Clin Microbiol. 2004 Feb;42(2):563-9. doi: 10.1128/JCM.42.2.563-569.2004.
In this multicenter evaluation, the VERSANT HCV RNA 3.0 Assay (bDNA) (Bayer Diagnostics, Tarrytown, N.Y.) was shown to have excellent reproducibility, linearity, and analytical sensitivity across specimen collection matrices (serum, EDTA, ACD-A), and hepatitis C virus (HCV) genotypes 1 to 6. The VERSANT HCV bDNA Assay has a reportable range of 615 to 7690000 (7.69 x 10(6)) IU/ml. The total coefficient of variation (CV) ranged from 32.4% at 615 IU/ml to 17% at 6.8 x 10(6) IU/ml. The assay was linear across the reportable range. Analytical specificity of 98.8% was determined by testing 999 specimens from volunteer blood donors. Evaluation of HCV genotypes using RNA transcripts of representative clones of 1a, 1b, 2a, 2b, 2c, 3a, 4a, 5a, and 6a and patient specimens showed that the largest difference between genotype 1, upon which the assay is standardized, and non-1 genotypes was within 1.5-fold. Testing of potentially interfering endogenous substances and exogenous substances and conditions found no interference in HCV-positive or HCV-negative specimens except for unconjugated bilirubin at concentrations of >or=20 mg/dl and protein at concentrations of >or=9 g/dl. Biological variability was estimated from 29 clinically stable individuals not on HCV therapy who were tested weekly over an 8-week period. The combined estimate of total (biologic plus assay) variability was 0.15 log(10) standard deviation (CV, 36.1%), a fold change of 2.6. Thus, the observed fold change between any two consecutive HCV RNA measures is expected to be less than 2.6-fold (equivalent to 0.41 log(10) IU/ml) 95% of the time in clinically stable individuals.
在这项多中心评估中,VERSANT HCV RNA 3.0检测法(分支DNA法)(拜耳诊断公司,纽约州塔里敦)在各种样本采集基质(血清、乙二胺四乙酸、酸性枸橼酸盐葡萄糖-A)以及丙型肝炎病毒(HCV)1至6型中均显示出出色的重复性、线性和分析灵敏度。VERSANT HCV分支DNA检测法的报告范围为615至7690000(7.69×10⁶)IU/ml。总变异系数(CV)在615 IU/ml时为32.4%,在6.8×10⁶ IU/ml时为17%。该检测法在报告范围内呈线性。通过检测999份来自志愿献血者的样本确定分析特异性为98.8%。使用1a、1b、2a、2b、2c、3a、4a、5a和6a代表性克隆的RNA转录本以及患者样本对HCV基因型进行评估,结果显示,该检测法所标准化的1型与非1型基因型之间的最大差异在1.5倍以内。对潜在干扰内源性物质和外源性物质及条件进行检测发现,除了浓度≥20 mg/dl的未结合胆红素和浓度≥9 g/dl的蛋白质外,在HCV阳性或HCV阴性样本中均未发现干扰。从29名未接受HCV治疗的临床稳定个体中估算生物学变异性,这些个体在8周内每周进行检测。总(生物学加检测)变异性的综合估算值为0.15 log₁₀标准差(CV,36.1%),变化倍数为2.6。因此,在临床稳定个体中,预计95%的情况下,任意两个连续的HCV RNA测量值之间观察到的变化倍数将小于2.6倍(相当于0.41 log₁₀ IU/ml)。