Detection, semiquantitation, and genetic variation in hepatitis C virus sequences amplified from the plasma of blood donors with elevated alanine aminotransferase.
Detection, semiquantitation, and genetic variation in hepatitis C virus sequences amplified from the plasma of blood donors with elevated alanine aminotransferase.
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从丙氨酸氨基转移酶升高的献血者血浆中扩增的丙型肝炎病毒序列的检测、半定量和遗传变异。
DOI：
10.1172/jci114882
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Vyas,GN
中科院分区：
文献类型：
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作者：
Ulrich,PP;Romeo,JM;Lane,PK;Kelly,I;Daniel,LJ;Vyas,GN
丙型肝炎病毒（HCV）是输血后非甲非乙型肝炎的主要病原体，以丙氨酸氨基转移酶（ALT）波动性升高和慢性肝病为特征。从ALT水平升高的无症状志愿献血者中获得101个不可检测的血浆单位，商业酶联免疫吸附试验在11个标本中检测到HCV抗体（抗-HCV）。使用我们开发的联合逆转录聚合酶链反应（RT-PCR）检测，HCVRNA检测0.6毫升血浆从8的11（73%）的抗-HCV阳性，但没有在90抗-HCV阴性标本。当分析2.4 ml血浆时，在剩余的3份抗HCV阳性标本中可检测到相对较低浓度的HCV RNA。病毒体的血浆浓度估计为10（2）至5 × 10（7）/ml。对来自三个标本的PCR扩增的HCV cDNA（210个碱基对）进行的直接测序显示，核苷酸水平的异质性在2.5%和8.6%之间，氨基酸水平的异质性小于4%。我们的研究结果表明，RT-PCR可以用2.4 ml血浆进行，提供了一种直接检测HCV RNA的检测方法，并证实了在表面健康的抗HCV阳性供体中存在HCV感染的无症状携带者状态。图片
Hepatitis C virus (HCV) is the predominant etiologic agent of posttransfusion non-A, non-B hepatitis, characterized by undulating elevation of alanine aminotransferase (ALT) and chronic liver disease. A commercial enzyme-linked immunosorbent assay detected antibodies to HCV (anti-HCV) in 11 specimens among 101 nontransfusable plasma units obtained from asymptomatic, volunteer blood donors with elevated levels' of ALT. Using a combined reverse-transcription polymerase chain reaction (RT-PCR) assay developed by us, HCV RNA was detected in 0.6 ml of plasma from 8 of 11 (73%) of the anti-HCV-positive but in none of the 90 anti-HCV-negative specimens. The relatively low concentration of HCV RNA could be detected in the remaining three anti-HCV-positive specimens when 2.4 ml of plasma was analyzed. The plasma concentration of virions was estimated to range from 10(2) to 5 x 10(7)/ml. Direct sequencing performed on the PCR-amplified HCV cDNAs (210 base pairs) from three specimens revealed heterogeneity between 2.5 and 8.6% at the nucleotide level and less than 4% at the amino acid level. Our findings demonstrate that RT-PCR can be performed with 2.4 ml of plasma, providing an assay for the direct detection of HCV RNA and confirming the existence of an asymptomatic carrier state for HCV infection in the apparently healthy anti-HCV-positive donors.Images