Sun R, Jayakar H, Yeh S, Mendoza M, Wisbeski M H, Iszczyszyn W, Dragon E A
Roche Molecular Systems, Inc., Pleasanton, CA 94566, USA.
Dev Biol (Basel). 2000;102:81-91.
The COBAS AmpliScreen Hepatitis C (HCV) Test, Version 2.0, which is designed for screening pools composed of samples from individual units of blood or plasma, employs a MultiPrep sample processing procedure that simultaneously extracts and concentrates HCV, HIV-1 and Hepatitis B virus particles from plasma. An HCV Internal Control (IC) RNA serves as an extraction and amplification control for each independently processed sample. Processed samples are amplified by RT-PCR using HCV-specific complementary primers and detected by hybridization of the amplified products to HCV- and IC-specific oligonucleotide probes. The analytical sensitivity of the test is 25 International Units (IU) of HCV per mL of pooled plasma; all HCV genotypes are detected with similar efficiency. The test detected HCV RNA 23 to 32 days prior to anti-HCV antibody seroconversion for four of the five seroconversion panels tested. The test had sufficient sensitivity to reproducibly detect a single infected unit containing 2.4 x 10(3) copies of HCV per mL in a pool with 23 uninfected units. COBAS AmpliScreen tests for HIV-1 and HBV now being validated by Roche Molecular Systems also incorporate the MultiPrep specimen processing method, thereby making it possible to use a single processed specimen to screen for all three viruses.
COBAS AmpliScreen丙型肝炎（HCV）检测试剂盒，版本2.0，专为筛查由单个血液或血浆样本组成的混合样本而设计，采用MultiPrep样本处理程序，可同时从血浆中提取并浓缩HCV、HIV-1和乙型肝炎病毒颗粒。HCV内部对照（IC）RNA用作每个独立处理样本的提取和扩增对照。处理后的样本使用HCV特异性互补引物通过RT-PCR进行扩增，并通过扩增产物与HCV和IC特异性寡核苷酸探针的杂交进行检测。该检测的分析灵敏度为每毫升混合血浆25国际单位（IU）的HCV；所有HCV基因型的检测效率相似。在测试的五个血清转化组中，有四个组在抗HCV抗体血清转化前23至32天检测到HCV RNA。该检测具有足够的灵敏度，可在含有23个未感染单位的混合样本中重复检测出每毫升含有2.4×10(3)个HCV拷贝的单个感染单位。罗氏分子系统公司目前正在验证的用于HIV-1和HBV的COBAS AmpliScreen检测也采用了MultiPrep样本处理方法，从而有可能使用单个处理后的样本筛查所有三种病毒。
