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用于检测子宫颈浸润癌中人乳头瘤病毒序列的Southern印迹分析与同位素和非同位素原位杂交的比较

Comparison of Southern blot analysis with isotopic and nonisotopic in situ hybridization for the detection of human papillomavirus sequences in invasive carcinoma of the uterine cervix.

作者信息

D'Amato L, Pilotti S, Rotola A, Di Luca D, Cassai E, Rilke F

机构信息

Division of Anatomical Pathology and Cytology, Istituto Nazionale Tumori, Milano, Italy.

出版信息

Mod Pathol. 1992 Mar;5(2):207-11.

PMID:1315439
Abstract

To compare the efficiency of hybridization methods for the detection of HPV genome, 22 cases of invasive squamous cell carcinoma of the uterine cervix were analyzed by Southern blot analysis and in situ hybridization carried out with 35S- and biotin-labeled probes. These cases contained from less than one to as many as 50 copies per cell of HPV 16 and 18 types. To increase the sensitivity of biotinylated probes, a silver enhancement procedure of the peroxidase reaction product was applied. Results showed that in situ hybridization performed with isotopic probes is as sensitive as Southern blot analysis and is more sensitive than that performed with biotin-labeled probe. However, the application of the silver enhancement procedure increases the percentage of HPV-positive cases from 27 to 50%.

摘要

为比较用于检测人乳头瘤病毒(HPV)基因组的杂交方法的效率,采用Southern印迹分析法以及用35S和生物素标记的探针进行原位杂交,对22例子宫颈浸润性鳞状细胞癌病例进行了分析。这些病例中,HPV 16型和18型每个细胞含有的拷贝数从少于1个到多达50个不等。为提高生物素化探针的敏感性,对过氧化物酶反应产物采用了银增强程序。结果显示,用同位素探针进行的原位杂交与Southern印迹分析法一样敏感,且比用生物素标记探针进行的原位杂交更敏感。然而,银增强程序的应用使HPV阳性病例的百分比从27%增至50%。

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