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常规处理的肝活检组织的丙型肝炎病毒聚合酶链反应

Hepatitis C virus-polymerase chain reaction of routinely processed liver biopsies.

作者信息

el-Batanony M H, Savage K, Jacobs R, el-Refaie A O, Squadrito G G, Brown D, Saleh S M, Raouf A A, Amer K M, Dusheiko G M

机构信息

University Department of Histopathology, Royal Free Hospital and School of Medicine, London, United Kingdom.

出版信息

J Med Virol. 1994 Aug;43(4):380-5. doi: 10.1002/jmv.1890430411.

Abstract

The aim of the study was to evaluate the specificity and sensitivity of detection of hepatitis C virus (HCV)-RNA in formalin-fixed paraffin-embedded (FFPE) liver biopsies by polymerase chain reaction (PCR). Routinely processed FFPE diagnostic needle liver biopsies as well as stored serum samples from 43 patients with liver disease were tested for HCV-RNA by reverse transcription-nested PCR using the same sets of primers and following strict anticontamination measures. Twenty-nine cases were positive and 14 were negative for serum HCV-RNA. Tissue HCV-RNA was detected in 17 out of the 29 serum HCV-RNA-positive cases but not in any of the 14 serum HCV-RNA-negative cases. Compared to serum-PCR, tissue-PCR was 100% specific, 58.6% sensitive, and 72% efficient. HCV-RNA was detected more frequently in biopsies stored for less than 1 year, than in those stored for more than 1 year (P = 0.046). In biopsies stored for up to 1 year detection of HCV-RNA by PCR was 81.8% sensitive and 90.9% efficient. Short (< 0.5 cm) liver biopsies were as sufficient for nucleic acid extraction and amplification as long (> 0.5 cm) ones. It is concluded that following strict anticontamination measures, HCV-RNA detection by PCR in routinely fixed, processed, and stored diagnostic liver biopsies provides a valuable adjunct to diagnosis of HCV infection. In this study, this option was free from contamination problems, even though routine batch histological processing schedules were used.

摘要

本研究旨在通过聚合酶链反应(PCR)评估在福尔马林固定石蜡包埋(FFPE)肝活检组织中检测丙型肝炎病毒(HCV)-RNA的特异性和敏感性。使用相同引物组并遵循严格的防污染措施,通过逆转录巢式PCR对43例肝病患者的常规处理的FFPE诊断性肝穿刺活检组织以及储存的血清样本进行HCV-RNA检测。血清HCV-RNA检测中,29例阳性,14例阴性。29例血清HCV-RNA阳性病例中有17例检测到组织HCV-RNA,而14例血清HCV-RNA阴性病例中均未检测到。与血清PCR相比,组织PCR的特异性为100%,敏感性为58.6%,效率为72%。储存时间少于1年的活检组织中HCV-RNA的检测频率高于储存时间超过1年的活检组织(P = 0.046)。在储存时间长达1年的活检组织中,PCR检测HCV-RNA的敏感性为81.8%,效率为90.9%。短(<0.5 cm)肝活检组织与长(>0.5 cm)肝活检组织在核酸提取和扩增方面同样充足。结论是,遵循严格的防污染措施,通过PCR在常规固定、处理和储存的诊断性肝活检组织中检测HCV-RNA可为HCV感染的诊断提供有价值的辅助手段。在本研究中,即使采用常规批量组织学处理方案,该方法也不存在污染问题。

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