Miyauchi I, Moriyama M, Zhang D Y, Abe K
Department of Pathology, National Institute of Infectious Diseases, Tokyo, Japan.
Pathol Int. 1998 Jun;48(6):428-32. doi: 10.1111/j.1440-1827.1998.tb03928.x.
Ligation-dependent polymerase chain reaction (LD-PCR) is a novel, simple and sensitive PCR assay that could be useful for the molecular diagnosis of various viral infections. This assay uses two capture probes for RNA isolation and two hemiprobes for the subsequent PCR. To investigate its usefulness in the field of pathological diagnosis, further study of this new PCR technology to detect hepatitis C virus (HCV) RNA was carried out in formalin-fixed, paraffin-embedded (FFPE) liver tissues and the results compared with those obtained by the reverse transcription (RT)-PCR method. We analyzed routinely processed FFPE liver specimens of surgical origin from 13 anti-HCV-seropositive Japanese patients with hepatocellular carcinoma (HCC). As a result, HCV-RNA was detected in FFPE liver specimens in 12 of the tested patients (92.3%) by the LD-PCR method without the use of reverse transcription. In contrast, HCV-RNA was detectable in eight out of 13 cases (61.5%) by the RT-PCR method. These results indicated that LD-PCR was useful for HCV-RNA detection in routinely processed FFPE liver specimens.
连接依赖型聚合酶链反应(LD-PCR)是一种新颖、简单且灵敏的聚合酶链反应检测方法,可用于多种病毒感染的分子诊断。该检测方法使用两个捕获探针进行RNA分离,并使用两个半探针进行后续的聚合酶链反应。为了研究其在病理诊断领域的实用性,我们在福尔马林固定、石蜡包埋(FFPE)的肝组织中对这种检测丙型肝炎病毒(HCV)RNA的新型聚合酶链反应技术进行了进一步研究,并将结果与通过逆转录(RT)-聚合酶链反应方法获得的结果进行了比较。我们分析了13例抗-HCV血清阳性的日本肝细胞癌(HCC)患者手术来源的常规处理的FFPE肝标本。结果,通过LD-PCR方法在12例受测患者(92.3%)的FFPE肝标本中检测到HCV-RNA,无需进行逆转录。相比之下,通过RT-PCR方法在13例病例中的8例(61.5%)中检测到HCV-RNA。这些结果表明,LD-PCR可用于常规处理的FFPE肝标本中HCV-RNA的检测。