Walker F, Bedel C, Dauge-Geffroy M C, Lehy T, Madelenat P, Potet F
Pathology Department, Bichat Hospital, Paris, France.
Diagn Mol Pathol. 1996 Jun;5(2):136-46. doi: 10.1097/00019606-199606000-00009.
The prevalence of genital human papillomavirus (HPV) infection was evaluated in 30 consecutive human immunodeficiency virus (HIV) + women by polymerase chain reaction (PCR)-in situ hybridization (ISH) on paraffin-embedded tissue sections and compared with that found with standard ISH. Biopsies were removed from normal or neoplastic areas in the cervix, vagina, and vulva, and ISH was performed with biotinylated or fluorescein isothiocyanate genomic DNA probes. One probe was used for HPV screening and others for HPV typing (types 6, 11, 16, 18, 31, and 33). Sequences were amplified by the "hot-start" PCR method and followed by standard ISH. Among the 30 HIV + women, 90% scored HPV + in one or several locations by PCR-ISH, whereas only 67% were positive by ISH. Oncogenic HPV types were found in 63% by PCR-ISH and in only 43% by ISH. The same HPV types detected by standard ISH were also recognized by PCR-ISH, but with the latter the signal was amplified. Moreover, some HPV types were found with PCR-ISH but not by ISH. We conclude that PCR-ISH is a valuable and sensitive method for specific detection of HPV.
通过对石蜡包埋组织切片进行聚合酶链反应(PCR)-原位杂交(ISH),评估了30例连续的人类免疫缺陷病毒(HIV)阳性女性的生殖道人乳头瘤病毒(HPV)感染率,并与标准ISH检测结果进行比较。从宫颈、阴道和外阴的正常或肿瘤区域获取活检组织,使用生物素化或异硫氰酸荧光素基因组DNA探针进行ISH。一种探针用于HPV筛查,其他探针用于HPV分型(6、11、16、18、31和33型)。通过“热启动”PCR方法扩增序列,然后进行标准ISH。在30例HIV阳性女性中,90%通过PCR-ISH在一个或多个部位检测到HPV阳性,而通过ISH检测仅67%为阳性。通过PCR-ISH检测到致癌性HPV类型的比例为63%,而通过ISH检测仅为43%。标准ISH检测到的相同HPV类型也能被PCR-ISH识别,但后者信号得到了放大。此外,通过PCR-ISH发现了一些ISH未检测到的HPV类型。我们得出结论,PCR-ISH是一种用于特异性检测HPV的有价值且敏感的方法。