Improved COBAS TaqMan hepatitis C virus test (version 2.0) for use with the high pure system: Enhanced genotype inclusivity and performance characteristics in a multisite study
Improved COBAS TaqMan hepatitis C virus test (version 2.0) for use with the high pure system: Enhanced genotype inclusivity and performance characteristics in a multisite study
复制标题
DOI：
10.1128/jcm.01320-07
复制
发表时间：
2007-11-01
影响因子：
9.4
通讯作者：
Sarrazin, C.
中科院分区：
文献类型：
--
作者：
Colucci, G.;Ferguson, J.;Sarrazin, C.
我们评价了COBAS TaqMan丙型肝炎病毒（HCV）检测试剂盒（2.0版）与High Pure系统（HCVHPS V2）的配合使用，这是一种新的修订版实时逆转录PCR检测试剂盒，旨在改进1.0版（HCVHPS V1）的基因型定量。在洗涤缓冲液和逆转录温度下进行修正。使用HCVHPS V2、HCVHPS V1和COBAS AMPLYCOR HCV MONITOR检测（版本2.0）（CAHCM），在三个不同的研究中心评价HCVHPS V2的基因型包容性。其中一个参与实验室还使用了全自动COBAS Ampliprep/COBAS TaqMan HCV检测。HCVHPS V2和CAHCM之间以及HCVHPS V2和HCVHPS V1之间的HCV RNA值的平均差异范围分别为-0.21到0.13 log和0.24到1.27 log，基因型2、3、4和5的差异> 0.5 log。对于HCV基因型1至6的NIBSC组，所有6种基因型的测得HCVHPS V2值均在标称值的0.25 log范围内。当检测基因型特异性临床HCV标本的系列稀释液时，该检测试剂盒的检出限为10 - 20 IU/ml，线性范围为25 IU/ml-3.91 x 10（8）IU/ml。用100份HCV血清阴性标本以及12份黄病毒科非HCV成员和22种其他微生物证明了100%的临床和分析特异性。这些数据表明，HCVHPS V2是一种用于定量所有6种HCV基因型的稳健且准确的检测方法，可用于监测所有临床HCV标本中的病毒载量。
We have evaluated the COBAS TaqMan hepatitis C virus (HCV) test (version 2.0) for use with the High Pure system (HCVHPS V2), a new, revised real-time reverse transcription-PCR assay developed to improve the genotype quantitation of version 1.0 (HCVHPS V1). Revisions were made in the wash buffer and in the reverse transcription temperature. The genotype inclusivity of HCVHPS V2 was evaluated at three different sites, using HCVHPS V2, HCVHPS V1, and the COBAS AMPLICOR HCV MONITOR test (version 2.0) (CAHCM). The fully automated COBAS Ampliprep/COBAS TaqMan HCV test was also used in one of the participating laboratories. The mean differences in HCV RNA values between HCVHPS V2 and CAHCM and between HCVHPS V2 and HCVHPS V1 ranged from -0.21 to 0.13 log and from 0.24 to 1.27 log, respectively, with > 0.5-log differences for genotypes 2, 3, 4, and 5. With a NIBSC panel of HCV genotypes 1 through 6, the measured HCVHPS V2 values were within 0.25 log of the nominal values for all 6 genotypes. When serial dilutions of genotype-specific clinical HCV specimens were tested, the assay showed a limit of detection between 10 and 20 IU/ml and a linear range of 25 IU/ml to 3.91 x 10(8) IU/ml. Clinical and analytical specificities of 100% were demonstrated with 100 HCV-seronegative specimens as well as with 12 non-HCV members of Flaviviridae and 22 additional microorganisms. These data indicate that HCVHPS V2 is a robust and accurate test for the quantitation of all six HCV genotypes and useful in monitoring viral load in all clinical HCV specimens.