Novel short-fragment PCR assay for highly sensitive broad-spectrum detection of anogenital human papillomaviruses
Novel short-fragment PCR assay for highly sensitive broad-spectrum detection of anogenital human papillomaviruses
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DOI：
10.1016/s0002-9440(10)65688-x
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发表时间：
1998-12-01
影响因子：
6
通讯作者：
Quint, W
中科院分区：
文献类型：
--
作者：
Kleter, B;van Doorn, LJ;Quint, W
为普遍检测人乳头瘤病毒(HPV)，开发了一套新的聚合酶链式反应(PCR)引物，命名为SPF1和SPF2，位于L1区。合成了一个短小的聚合酶链式反应片段(SPF)，其pH值仅为65。在一个基于微量滴度的杂交系统中，使用混合的寡核苷酸探针检测到SPF扩增。SPF系统可以检测到至少43种不同的HPV基因类型。在三个不同的患者组中评估了新的SPF系统的临床性能。1)对534例宫颈刮片的检测结果显示，细胞学正常的宫颈刮片中有447例(83.7%)的阳性率明显高于通用的Gp5+/6+(P<；0.001)。(2)SPF法检测304例细胞学异常刮片中的299例(98.4%)HPVDNA。3)SPF系统在184例福尔马林固定、石蜡包埋的宫颈癌标本中100%检测到HPVDNA。总之，新的SPF系统允许在不同的临床材料中普遍和高灵敏度地检测HPV DNA，并可能改进这种重要病毒的分子诊断和流行病学。
A novel set of polymerase chain reaction (PCR) primers, designated SPF1 and SPF2 and located in the L1 region, was developed for universal detection of human papillomavirus (HPV). A short PCR fragment (SPF) of only 65 ph was synthesized. SPF amplimers were detected in a microtiter-based hybridization system, using a mixture of oligonucleotide probes. The SPF system allowed detection of at least 43 different HPV genotypes. The clinical performance of the novel SPF system was assessed in three different patient groups. 1) Analysis of 534 cervical scrapes, obtained from treated patients, showed that the detection rate in 447 (83.7%) scrapes with normal cytology was significantly higher using the SPF system as compared with the universal primer set GP5 +/6+ (P < 0.001). 2) The SPF assay detected HPV DNA in 299 (98.4%) of 304 scrapes with cytological dyskaryosis. 3) The SPF system detected HPV DNA in 100% of 184 formalin-fixed, paraffin-embedded cervical carcinoma specimens. In conclusion, the novel SPF system permitted universal and highly sensitive detection of HPV DNA in diverse clinical materials and may improve the molecular diagnosis and epidemiology of this important virus.