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[应用聚合酶链反应和原位杂交技术检测子宫颈病变中的人乳头瘤病毒DNA]

[Detection of HPV DNA in the uterine cervical lesions by polymerase chain reaction and in situ hybridization].

作者信息

Kyo S, Inoue M, Nakazawa A, Ogawa H, Ueda G, Tanizawa O

机构信息

Department of Obstetrics and Gynecology, Osaka University School of Medicine.

出版信息

Nihon Sanka Fujinka Gakkai Zasshi. 1991 May;43(5):541-8.

PMID:1647430
Abstract

Human papillomavirus (HPV) is found in close association with carcinogenesis of the uterine cervix. We applied a new in vitro gene amplification technology, the polymerase chain reaction (PCR) to detect HPV 16 and 18 in cervical exfoliated cells. HPV infections were detected in 5 (16%) of 31 women with no pathological lesions of the uterine cervix (normal), 16 (24%) of 67 with cervical intraepithelial neoplasia (CIN) and 6 (38%) of 16 with invasive cervical cancer. Moreover, 10% formalin-fixed and paraffin-embedded tissue sections were prepared from the uterine cervix of these 27 women with PCR-proven HPV infection and were examined for the histological localization of HPV-DNA by in situ hybridization with biotin-labeled DNA probes of HPV types 6/11, 16/18 and 31/33/35. HPV-DNA type 16/18 was detected in 3 of 5 normal women, 2 of 4 CINs I, 2 of 3 CINs II, 6 of 9 CINs III and 6 of 6 invasive cervical cancers. HPV-DNA type 6/11 was detected in 6 of 6 condylomas. Viral DNA sequence was detected in the superficial cells of CIN I and II, and it was distributed through entire thickness layer of undifferentiated cells derived from CIN III and squamous cell carcinoma. In addition, the staining intensity became weak as the lesion progressed. These differences between lesions might be due to the difference in the viral form in the nuclei, ie whether an episomal or integrated form. Thus, an in situ hybridization technique with a biotin-labeled DNA probe as well as the PCR method is useful for the detection of HPV in clinical samples.

摘要

人乳头瘤病毒(HPV)与子宫颈癌的发生密切相关。我们应用一种新的体外基因扩增技术——聚合酶链反应(PCR)来检测宫颈脱落细胞中的HPV 16和18。在31名子宫颈无病理病变(正常)的女性中,有5名(16%)检测到HPV感染;在67名患有宫颈上皮内瘤变(CIN)的女性中,有16名(24%)检测到HPV感染;在16名浸润性宫颈癌患者中,有6名(38%)检测到HPV感染。此外,从这27名经PCR证实感染HPV的女性的子宫颈制备了10%福尔马林固定石蜡包埋组织切片,并用HPV 6/11、16/18和31/33/35型生物素标记DNA探针进行原位杂交,检测HPV-DNA的组织学定位。在5名正常女性中的3名、4名CIN I中的2名、3名CIN II中的2名、9名CIN III中的6名以及6名浸润性宫颈癌患者中的6名中检测到HPV-DNA 16/18型。在6名尖锐湿疣患者中的6名检测到HPV-DNA 6/11型。在CIN I和II的表层细胞中检测到病毒DNA序列,并且它分布在源自CIN III和鳞状细胞癌的未分化细胞的全层。此外,随着病变进展,染色强度变弱。病变之间的这些差异可能是由于细胞核中病毒形式的差异,即是否为游离型或整合型。因此,生物素标记DNA探针原位杂交技术以及PCR方法对于临床样本中HPV的检测是有用的。

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