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宫颈癌石蜡包埋切片中特定人乳头瘤病毒类型的检测

Detection of specific human papillomavirus types in paraffin-embedded sections of cervical carcinomas.

作者信息

Bryan Janine T, Taddeo Frank, Skulsky Deemarie, Jansen Kathrin U, Frain Barbara M, Qadadri Brahim, Brown Darron R

机构信息

Merck Research Laboratories, West Point, Pennsylvania, USA.

出版信息

J Med Virol. 2006 Jan;78(1):117-24. doi: 10.1002/jmv.20512.

Abstract

Human papillomaviruses (HPV) are the causative agents of most cervical carcinomas. A complete understanding of the HPV types that cause cervical carcinoma is needed as vaccines are designed. Fresh tissues are not always available for such studies. We therefore sought to determine the feasibility of HPV studies using formalin-fixed, paraffin-embedded sections of 56 cervical carcinomas, correlating typing information with the pathology and physical state of the HPV sequences within cells. Sections from each specimen were used to extract and purify DNA. Specific HPV types were identified using a PCR/reverse blot strip assay. Tyramide signal-amplified, fluorescent DNA in situ hybridization (FISH) was used to localize HPV within cells. Human beta-globin sequences were amplified in DNA from all specimens. HPV sequences from oncogenic types were identified in 52 of 56 (92.9%) by PCR/reverse blot strip assay, and in one additional case using an HPV 16 multiplex PCR assay. HPV 16 was the most commonly detected type, present in most cases as a solitary isolate. Thirty- five of 42 HPV 16 or HPV 18 PCR-positive specimens were also positive in the FISH assay, in most cases in a pattern consistent with viral integration. We conclude that HPV typing from formalin-fixed, paraffin-embedded sections of cervical carcinomas is possible, with a sensitivity that is similar to that found in studies using fresh tissue.

摘要

人乳头瘤病毒(HPV)是大多数宫颈癌的致病因子。由于疫苗的设计需要,因此有必要全面了解导致宫颈癌的HPV类型。此类研究并非总能获取新鲜组织。因此,我们试图确定利用56例宫颈癌的福尔马林固定、石蜡包埋切片进行HPV研究的可行性,将分型信息与细胞内HPV序列的病理及物理状态相关联。每个标本的切片用于提取和纯化DNA。使用PCR/反向印迹条带分析法鉴定特定的HPV类型。采用酪胺信号放大荧光DNA原位杂交(FISH)法在细胞内定位HPV。在所有标本的DNA中均扩增出人β-珠蛋白序列。通过PCR/反向印迹条带分析法在56例中的52例(92.9%)中鉴定出致癌型HPV序列,另外1例采用HPV 16多重PCR分析法鉴定。HPV 16是最常检测到的类型,在大多数情况下以单一分离株形式存在。42例HPV 16或HPV 18 PCR阳性标本中有35例在FISH检测中也呈阳性,在大多数情况下其模式与病毒整合一致。我们得出结论,从宫颈癌福尔马林固定、石蜡包埋切片中进行HPV分型是可行的,其灵敏度与使用新鲜组织的研究相似。

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