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短片段聚合酶链反应反向杂交线性探针检测法用于检测多种人乳头瘤病毒类型并进行基因分型。临床评估与随访。

Short fragment polymerase chain reaction reverse hybridization line probe assay to detect and genotype a broad spectrum of human papillomavirus types. Clinical evaluation and follow-up.

作者信息

Melchers W J, Bakkers J M, Wang J, de Wilde P C, Boonstra H, Quint W G, Hanselaar A G

机构信息

Department of Medical Microbiology, University of Nijmegen, Nijmegen Delft, The Netherlands.

出版信息

Am J Pathol. 1999 Nov;155(5):1473-8. doi: 10.1016/S0002-9440(10)65462-4.

Abstract

The purpose of this study was to detect and genotype 16 different human papilloma virus (HPV) types simultaneously using a short fragment polymerase chain reaction (SPF) hybridization line probe assay (LiPA). 152 women who were referred to the gynecologist because of abnormal cervical smear underwent colposcopic examination and repeat cervical smear. In addition, the cervical scrapes were analyzed for the presence of HPV by a novel general HPV polymerase chain reaction assay followed by a single reaction genotyping assay allowing for a simultaneous detection and identification of 16 different HPV types. HPV DNA was detected in 38% of normal follow-up cervical scrapes, 51% of scrapes with atypical squamous cells of undetermined significance, 78% of scrapes with mild dysplasia (low grade squamous intraepithelial lesions), 86% of scrapes with moderate dysplasia (high grade squamous intraepithelial lesions), and in 88% of scrapes with severe dysplasia and carcinoma in situ. One case of invasive squamous cell carcinoma was positive for HPV 16. Overall, a single HPV type was detected in 56% of HPV positive scrapes, with HPV 16 being the most common and accounting for 45% of all single infections. Forty-four percent of the positive scrapes contained multiple HPV types, of which double infections prevailed. Follow-up results proved the reproducibility and reliability of SPF HPV LiPA. In conclusion, we have used and evaluated the SPF-HPV-LiPA system for the detection and genotyping of HPV infections. The combined detection-typing method proved to be sensitive, specific, simple, and fast, making mass screening of cervical scrapes accessible for routine practice and facilitating individual patient management.

摘要

本研究的目的是使用短片段聚合酶链反应(SPF)杂交线性探针分析(LiPA)同时检测16种不同的人乳头瘤病毒(HPV)类型并进行基因分型。152名因宫颈涂片异常而转诊至妇科医生处的女性接受了阴道镜检查和重复宫颈涂片检查。此外,通过一种新型的通用HPV聚合酶链反应分析方法对宫颈刮片进行HPV检测,随后进行单反应基因分型分析,可同时检测和鉴定16种不同的HPV类型。在正常随访的宫颈刮片中,38%检测到HPV DNA;意义不明确的非典型鳞状细胞刮片中,51%检测到HPV DNA;轻度发育异常(低级别鳞状上皮内病变)刮片中,78%检测到HPV DNA;中度发育异常(高级别鳞状上皮内病变)刮片中,86%检测到HPV DNA;重度发育异常和原位癌刮片中,88%检测到HPV DNA。1例浸润性鳞状细胞癌患者的HPV 16呈阳性。总体而言,56%的HPV阳性刮片中检测到单一HPV类型,其中HPV 16最为常见,占所有单一感染的45%。44%的阳性刮片中含有多种HPV类型,其中双重感染最为普遍。随访结果证明了SPF HPV LiPA的可重复性和可靠性。总之,我们使用并评估了SPF-HPV-LiPA系统用于HPV感染的检测和基因分型。这种联合检测分型方法被证明是敏感、特异、简单且快速的,使得宫颈刮片的大规模筛查可用于常规实践,并便于对个体患者进行管理。

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