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丙型肝炎患者唾液和尿液中丙型肝炎病毒基因组的检测:单轮聚合酶链反应法检测HCV基因组的实用性

Demonstration of hepatitis C virus genome in saliva and urine of patients with type C hepatitis: usefulness of the single round polymerase chain reaction method for detection of the HCV genome.

作者信息

Numata N, Ohori H, Hayakawa Y, Saitoh Y, Tsunoda A, Kanno A

机构信息

Department of Microbiology, Sendai Municipal Institute of Public Health, Japan.

出版信息

J Med Virol. 1993 Oct;41(2):120-8. doi: 10.1002/jmv.1890410207.

Abstract

The possible transmission routes of hepatitis C virus (HCV) in patients without overt parenteral exposure (sporadic or community acquired form) were examined. Saliva and urine specimens obtained from type C hepatitis patients, whose sera were positive for the HCV genome, were examined by reverse transcription and polymerase chain reaction (RT-PCR). By analyzing the factors that influenced the detection of the HCV genome by PCR, we developed a single round method which enabled semiquantitative detection with higher sensitivity than that obtained with nested PCR. Single round PCR revealed that 34.8% (8 of 23) of saliva and 56.5% (13 of 23) of urine specimens from patients with type C hepatitis contained the HCV genome. The amounts of HCV genome in saliva and urine specimens correlated with those in serum. The relative amounts of HCV genome in serum, saliva, and urine from a chronic type C hepatitis patient were determined by comparing the reciprocal of the smallest volume of the specimens in which the PCR products were visualized in agarose gels (PCR units/ml), and the values were 1 x 10(5), 5 x 10(1), and 3 x 10(1) PCR units/ml for serum, saliva, and urine specimens, respectively.

摘要

对无明显肠道外暴露(散发或社区获得性形式)的丙型肝炎病毒(HCV)患者可能的传播途径进行了研究。从血清HCV基因组呈阳性的丙型肝炎患者获取唾液和尿液标本,采用逆转录聚合酶链反应(RT-PCR)进行检测。通过分析影响PCR检测HCV基因组的因素,我们开发了一种单轮方法,该方法能够进行半定量检测,且灵敏度高于巢式PCR。单轮PCR显示,丙型肝炎患者的唾液标本中有34.8%(23份中的8份)、尿液标本中有56.5%(23份中的13份)含有HCV基因组。唾液和尿液标本中HCV基因组的含量与血清中的含量相关。通过比较在琼脂糖凝胶中可见PCR产物的标本最小体积的倒数(PCR单位/毫升),测定了一名慢性丙型肝炎患者血清、唾液和尿液中HCV基因组的相对含量,血清、唾液和尿液标本的值分别为1×10⁵、5×10¹和3×10¹PCR单位/毫升。

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