Gómez M A, Drut R, Lojo M M, Drut R M
Cátedra de Genética Microbiana, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Argentina.
Medicina (B Aires). 1995;55(3):213-7.
We examined the presence and subtypes of human papillomavirus (HPV) in 20 paraffin-embedded samples (from 12 patients) of juvenile laryngeal papillomatosis using the polymerase chain reaction (PCR). The biopsies had been stored for months to 12 years. Due to the great genetic variability of HPV, we selected a conservative sequence of the viral genome (L1 region) to identify the vast majority of the subtypes. Positive results were obtained by one-step PCR amplification with the MY09-11 consensus primers (L1 region) in only 10 of the cases. After a two-step amplification (nested-PCR) with GP5-6 primers the 20 samples proved to be positive demonstrating the higher sensitivity of this method. In order to amplify a highly variable region of the genome (E6), specific primers for HPV types 6 and 11 (H6/11 L1-R2) were used. 7/12 patients were positive for this subtype. Since more that one subtype has been reported in the same sample, the presence of HPV 6-11 sequences does not exclude that other subtypes might be involved. The results of this study show that: 1) HPV is present in JLP. 2) The most frequent HPV subtype involved was from the 6-11 group. 3) PCR can be successfully used in archived tissue routinely processed in a laboratory of pathology.
我们使用聚合酶链反应(PCR)检测了20份(来自12名患者)青少年喉乳头状瘤石蜡包埋样本中的人乳头瘤病毒(HPV)的存在情况及其亚型。这些活检样本已保存数月至12年。由于HPV的基因变异性很大,我们选择了病毒基因组的保守序列(L1区域)来鉴定绝大多数亚型。仅在10例病例中通过使用MY09 - 11共有引物(L1区域)进行一步PCR扩增获得了阳性结果。在用GP5 - 6引物进行两步扩增(巢式PCR)后,20个样本均被证明为阳性,表明该方法具有更高的灵敏度。为了扩增基因组的高变区(E6),使用了针对HPV 6型和11型的特异性引物(H6/11 L1 - R2)。12名患者中有7名该亚型呈阳性。由于在同一样本中已报道有不止一种亚型,HPV 6 - 11序列的存在并不排除可能涉及其他亚型。本研究结果表明:1)HPV存在于青少年喉乳头状瘤中。2)最常见的HPV亚型属于6 - 11组。3)PCR可成功用于病理实验室常规处理的存档组织。