Ishi K, Kina K, Koyatsu J, Kawashima T, Suzuki F, Saito A, Kubota T, Igari J
Department of Clinical Laboratory, Juntendo University Urayasu Hospital, Urayasu.
Rinsho Byori. 1997 Jan;45(1):82-6.
The causal association of human papilloma virus(HPV) with cervical cancer has been supported by multiple lines of evidence. Therefore, in the case of dysplasia, the presence of HPV-DNA should be detected and its subtypes identified. This is important in the determination of the prognosis for cervical disease. We reported a study in which the localization and types of HPV in cervical diseases was identified by in situ polymerase chain reaction(PCR), using biotin-labelled DNA probes. The in situ PCR, used by us was modified of Nuovo's method. We used biopsy materials of 18 CIN and 9 SCC cases(total 27 cases), all of which had been detected HPV-DNA by Southern blot hybridization, but not detected by in situ hybridization. A positive intranuclear reaction was detected in 13 of 18 CIN cases and 6 of 9 SCC cases(total 19 positive cases). Molecular biological techniques are the most reliable methods for detecting specific tumor genes and virus DNA. In situ hybridization has the advantage of enabling recognition of the cellular localization of the DNA in histologic specimens, but its sensitivity in inferior to the other techniques such as Southern blot, Dot blot and PCR. In situ PCR method possesses the advantages of both PCR and in situ hybridization in being highly sensitive and enabling visualization of the cellular localization of the DNA. In our present study, we succeeded to detect HPV-DNA in cervical biopsies of CIN and SCC cases by the in situ PCR.
人乳头瘤病毒(HPV)与宫颈癌之间的因果关联已得到多方面证据的支持。因此,对于发育异常的情况,应检测HPV-DNA的存在并确定其亚型。这对于判断宫颈疾病的预后很重要。我们报告了一项研究,其中通过使用生物素标记的DNA探针的原位聚合酶链反应(PCR)来确定宫颈疾病中HPV的定位和类型。我们使用的原位PCR是对Nuovo方法的改进。我们使用了18例CIN和9例SCC病例(共27例)的活检材料,所有这些病例均已通过Southern印迹杂交检测到HPV-DNA,但原位杂交未检测到。在18例CIN病例中有13例以及9例SCC病例中有6例(共19例阳性病例)检测到阳性核内反应。分子生物学技术是检测特定肿瘤基因和病毒DNA最可靠的方法。原位杂交的优点是能够在组织学标本中识别DNA的细胞定位,但其敏感性低于Southern印迹、斑点印迹和PCR等其他技术。原位PCR方法兼具PCR和原位杂交的优点,既高度敏感又能使DNA的细胞定位可视化。在我们目前的研究中,我们通过原位PCR成功地在CIN和SCC病例的宫颈活检中检测到了HPV-DNA。