Rady P L, Arany I, Hughes T K, Tyring S K
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston 77555-1019, USA.
J Virol Methods. 1995 Jun;53(2-3):245-54. doi: 10.1016/0166-0934(95)00029-t.
Consensus degenerate primers specific for the L1 and E1 regions were used to amplify anogenital HPV-DNA fragments. The mixture of the viral fragments was then directly sequenced with HPV-6, -11, -16, -18 and -33 type-specific computer-designed oligonucleotides as sequencing primers. The linking of a consensus primer-generated PCR amplification with type-specific primer-mediated direct sequencing and computer data bank analysis provided precise, more objective viral detection in the simultaneous presence of different HPV types. The method was successfully adapted for viral typing in clinical lesions which simultaneously contained different anogenital HPV sequences.
用于L1和E1区域的共有简并引物被用于扩增肛门生殖器HPV-DNA片段。然后，使用HPV-6、-11、-16、-18和-33型特异性计算机设计的寡核苷酸作为测序引物，对病毒片段混合物进行直接测序。将共有引物介导的PCR扩增与型特异性引物介导的直接测序及计算机数据库分析相结合，能够在同时存在不同HPV类型的情况下进行精确、更客观的病毒检测。该方法已成功应用于同时包含不同肛门生殖器HPV序列的临床病变的病毒分型。
