Abundance of Multiple High-Risk Human Papillomavirus (HPV) Infections Found in Cervical Cells Analyzed by Use of an Ultrasensitive HPV Genotyping Assay
Abundance of Multiple High-Risk Human Papillomavirus (HPV) Infections Found in Cervical Cells Analyzed by Use of an Ultrasensitive HPV Genotyping Assay
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DOI：
10.1128/jcm.00991-09
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发表时间：
2010-01-01
影响因子：
9.4
通讯作者：
Gheit, Tarik
中科院分区：
文献类型：
--
作者：
Schmitt, Markus;Dondog, Bolormaa;Gheit, Tarik
PCR方法能够检测感染肛门生殖道的各种各样的人乳头瘤病毒（HPV）基因型。然而，使用共有引物、通用引物以及在较小程度上使用广谱引物的PCR可能不能充分代表HPV的真实患病率，尤其是多重感染的真实患病率。我们比较了735例选定的宫颈刮片样本的HPV阳性率，其中使用引物BSGP 5+和BSGP 6+的广谱PCR（BS-PCR）与已建立的基于微珠的多重HPV基因分型（MPG）检测法相结合，使用类型特异性引物的多重PCR（TS-PCR）与新开发的MPG检测法相结合。虽然用于BS-PCR的引物位于HPV基因组的L1区域内，但用于TS-PCR的引物靶向E7基因。BS-PCR和TS-PCR对19种HPV类型的总阳性率分别为60.9%和72.2%，两种检测方法分别在34.8%和58.0%的标本中发现了多重感染。两种HPV检测方法均允许对HPV类型进行半定量检测，并在66.6%的多重感染中鉴定出相同的主要HPV类型。总之，TS-PCR-MPG检测显著增加了HPV DNA的检出率和检出的多种HPV类型感染的数量，并证明常规HPV扩增方法可能严重低估了肛门生殖道低拷贝数HPV感染的患病率，特别是在多种感染的情况下。因此，PCR-TS-MPG似乎非常适合于分析宫颈癌发展中多重感染的意义，以及研究HPV的自然史和潜伏期。
PCR methods enable the detection of a large variety of human papillomavirus (HPV) genotypes that infect the anogenital tract. However, PCR with consensus primers, general primers, and, to a lesser extent, broad-spectrum primers may underrepresent the true prevalence of HPV, especially the true prevalence of multiple infections. We compared the rate of HPV positivity determined by a broad-spectrum PCR with primers BSGP5+ and BSGP6+ (BS-PCR) coupled to an established bead-based multiplex HPV genotyping (MPG) assay with the rate of HPV positivity determined by a multiplex PCR with type-specific primers (TS-PCR) coupled to a newly developed MPG assay for 735 selected cervical scraping samples. While the primers used for the BS-PCR are located within the L1 region of the HPV genome, the primers used for the TS-PCR target the E7 gene. The overall rates of positivity for the 19 HPV types included in both assays were 60.9% and 72.2% by the BS-PCR and the TS-PCR, respectively, and the two assays found multiple infections in 34.8% and 58.0% of the specimens, respectively. Both HPV detection assays allowed the semiquantitative detection of HPV types and identified the same dominant HPV type in 66.6% of the multiple infections. In conclusion, the TS-PCR-MPG assay significantly increased the rate of detection of HPV DNA and the number of infections with multiple HPV types detected and demonstrated that the prevalence of low-copy-number HPV infections in the anogenital tract may be strongly underestimated by conventional HPV amplification methods, especially in cases of multiple infections. As a consequence, PCR-TS-MPG appears to be highly suited for analysis of the significance of multiple infections in the development of cervical cancer and for the study the natural history and the latency of HPV.