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用于特异性检测泰国肝细胞癌和胆管癌患者华支睾吸虫DNA的TaqMan实时荧光定量PCR检测法

TaqMan real-time PCR assay for specific detection of Opisthorchis viverrini DNA in Thai patients with hepatocellular carcinoma and cholangiocarcinoma.

作者信息

Suksumek Nithikoon, Leelawat Kawin, Leelawat Surang, Russell Bruce, Lek-Uthai Usa

机构信息

Division of Molecular Laboratory, Vachira Hospital, Bangkok 10400, Thailand.

出版信息

Exp Parasitol. 2008 Jun;119(2):217-24. doi: 10.1016/j.exppara.2008.01.018. Epub 2008 Feb 9.

Abstract

The aim of this study was to develop TaqMan real-time PCR assay that detected Opisthorchis viverrini DNA from 18 normal and 18 tumor tissue specimens from Thai patients with hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA), who underwent liver resection from October 2005 to May 2006. Control liver specimens were seven non-primary liver cancers. A conserved probe representing 100% sequence homology was used as a reference for O. viverrini-specific probe. Five of six tumors (83%) and all six normal tissues from CCA group; and seven of twelve tumors (58%) and ten of twelve normal tissues (83%) from HCC group were found to have O. viverrini DNA. The O. viverrini DNA detection among HCC and CCA patients were not associated (p=0.193; 90%CI). This RT-PCR will be a useful tool for investigating the relationship between cancer type and presence of the parasite and also for conducting epidemiological surveys.

摘要

本研究的目的是开发一种TaqMan实时荧光定量PCR检测方法,用于检测2005年10月至2006年5月期间接受肝切除术的泰国肝细胞癌(HCC)和胆管癌(CCA)患者的18份正常组织标本和18份肿瘤组织标本中的华支睾吸虫DNA。对照肝脏标本为7例非原发性肝癌。使用一个具有100%序列同源性的保守探针作为华支睾吸虫特异性探针的参考。CCA组6个肿瘤中的5个(83%)以及所有6个正常组织;HCC组12个肿瘤中的7个(58%)以及12个正常组织中的10个(83%)被发现含有华支睾吸虫DNA。HCC和CCA患者中华支睾吸虫DNA的检测结果无相关性(p = 0.193;90%置信区间)。这种逆转录PCR将成为研究癌症类型与寄生虫存在之间关系以及进行流行病学调查的有用工具。

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