Shibata D, Fu Y S, Gupta J W, Shah K V, Arnheim N, Martin W J
Department of Pathology, Los Angeles County-University of Southern California Medical Center.
Lab Invest. 1988 Oct;59(4):555-9.
Detection of human papilloma virus (HPV) types 16 and 18 in formalin-fixed, paraffin-embedded tissue by a new in vitro DNA amplification method, the polymerase chain reaction, was compared with detection with genomic DNA probes using in situ hybridization. The polymerase chain reaction replicates exponentially HPV DNA sequences present in a single 5- to 10-micron paraffin-embedded tissue section. The amplified sequences are detected with a DNA hybridization probe in a dot blot assay. The HPV polymerase chain reaction was able to detect on the average less than one HPV genome/cell as determined by tests of paraffin sections of cell pellets with known HPV genomic content. Cervical sections from 21 patients with HPV types 16, 18, or 31 as determined by in situ DNA hybridization were analyzed by the polymerase chain reaction. No disagreements between the two methods were detected. The sections comprising normal and dysplastic epithelium were further analyzed by the HPV polymerase chain reaction. The presence of virus correlated with the presence of dysplasia in the sections, though 3 of 10 normal sections contained HPV, and 1 of 21 sections with dysplasia lacked HPV 16 or 18. The polymerase chain reaction can specifically detect HPV 16 or 18 with high sensitivity from paraffin-embedded tissues.
采用一种新的体外DNA扩增方法——聚合酶链反应,对福尔马林固定、石蜡包埋组织中的人乳头瘤病毒16型和18型进行检测,并与使用基因组DNA探针的原位杂交检测方法进行比较。聚合酶链反应能使存在于单个5至10微米石蜡包埋组织切片中的人乳头瘤病毒DNA序列呈指数级复制。扩增后的序列通过斑点杂交检测法用DNA杂交探针进行检测。通过对已知人乳头瘤病毒基因组含量的细胞沉淀石蜡切片进行检测确定,人乳头瘤病毒聚合酶链反应平均能够检测到每个细胞中少于一个人乳头瘤病毒基因组。对21例经原位DNA杂交确定感染人乳头瘤病毒16型、18型或31型的患者的宫颈切片进行聚合酶链反应分析。未检测到两种方法之间存在不一致情况。对包含正常和发育异常上皮的切片进一步进行人乳头瘤病毒聚合酶链反应分析。切片中病毒的存在与发育异常的存在相关,不过10例正常切片中有3例含有病毒,21例发育异常切片中有1例缺乏人乳头瘤病毒16型或18型。聚合酶链反应能够从石蜡包埋组织中特异性地高灵敏度检测到人乳头瘤病毒16型或18型。