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Does human papillomavirus play a role in the development of bladder transitional cell carcinoma? A comparison of PCR and immunohistochemical analysis

机译:人乳头瘤病毒是否在膀胱移行细胞癌的发展中起作用? PCR和免疫组化分析的比较

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摘要

>Aim: To investigate the role of human papillomavirus (HPV) in the development of bladder transitional cell carcinoma (TCC).>Methods: Seventy eight paraffin wax embedded TCC samples were tested for the presence of HPV by two methods. First, immunohistochemistry was carried out using a polyclonal antibody capable of detecting the capsid protein of all known papillomaviruses. The second method was a consensus GP5+/6+ primer mediated polymerase chain reaction (PCR) technique, with the products analysed by both agarose gel electrophoresis and an enzyme immunoassay using type specific oligonucleotide probes for 10 different mucosal genotypes. To exclude false negative results because of the poor quality of DNA extracted from paraffin wax embedded samples, the series was extended to include 20 further blocks for which the corresponding snap frozen unfixed tissue was available.>Results: The two methods produced contrasting results, with 47 of the 78 samples positive for HPV antigen and none positive for HPV DNA. HPV DNA was not detected in the 20 additional paraffin wax embedded TCCs or in the 20 paired unfixed samples. In contrast, HPV DNA was amplified by PCR from all six of the paraffin wax embedded cervical carcinoma and anogenital wart control samples.>Conclusion: The disparity between the two sets of results is probably caused by false positives resulting from the non-specificity of the polyclonal antibody used for immunohistochemistry. These results suggest that HPV is unlikely to play an aetiological role in the development of bladder TCC.
机译:>目标:研究人乳头瘤病毒(HPV)在膀胱移行细胞癌(TCC)发生中的作用。>方法:测试了78份石蜡包埋的TCC样品HPV的存在有两种方法。首先,使用能够检测所有已知乳头瘤病毒的衣壳蛋白的多克隆抗体进行免疫组织化学。第二种方法是通用的GP5 + / 6 +引物介导的聚合酶链反应(PCR)技术,通过琼脂糖凝胶电泳和酶免疫法对产物进行分析,使用10种不同粘膜基因型的类型特异性寡核苷酸探针进行分析。为了排除由于从石蜡包埋的样品中提取的DNA质量差而导致的假阴性结果,该系列扩展到包括20个可用的相应速冻未固定组织的其他块。>结果:这些方法产生了相反的结果,在78个样本中有47个样本对HPV抗原呈阳性,而对HPV DNA没有阳性。在另外20个石蜡包埋的TCC中或在20对未固定的样品中未检测到HPV DNA。相比之下,通过PCR从所有六个石蜡包埋的宫颈癌和生殖器疣对照样品中扩增HPV DNA。>结论:两组结果之间的差异可能是由于以下原因导致的假阳性结果用于免疫组织化学的多克隆抗体的非特异性。这些结果表明HPV不太可能在膀胱TCC的发展中起病因作用。

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