人乳头瘤病毒DNA的原位杂交检测

DETECTION OF HUMAN PAPILLOMAVIRUS DNA BY IN SITU HYBRIDIZATION

①目的探讨原位杂交检测人乳头瘤病毒（ＨＰＶ）对尖锐湿疣的病因诊断意义及原位杂交的方法学。②方法用地高辛标记原位杂交技术，对４５例临床疑为尖锐湿疣的组织进行了ＨＰＶ６Ｂ／１１ＤＮＡ检测。为减轻非特异性背景染色并获得最大杂交敏感性，对消化酶、变性及杂交温度和时间、洗涤液、显色剂等原位杂交方法进行了研究。③结果３２例病理诊断为尖锐湿疣者均呈阳性，４例病理疑诊为尖锐湿疣者呈阴性，９例病理诊断为假性湿疣者均为阴性。

Objective To investigate the significance of human papillomavirus(HPV) detection by in situ hybridization in the diagnosis of condylomata acuminata and to study the methodology of in situ hybridization technique. Method Digoxigenin labelled in situ hybridization technique was used to detect HPV6B/11 DNA in 45 cases of tissue specimens which were suspected clinically as condylomata acuminata. In order to reduce the non specific background staining and obtain the maximal sensitivity, different concentrations of digestive enzyme, different temperature and time for denaturation and hybridization, stringency washes and reagents for detection were studied. Result 32 out of 45 cases showed positive results in both HPV in situ hybridization and histologic study. Four cases which were suspected for condylomata acuminata under microscope and 9 cases which were diagnosed histopathologically as psudo condylomata were all HPV negtive. Conclusion HPV in situ hybridization is a better method for the diagnosis of condylomata acluminata.