Potential of the in situ polymerase chain reaction in diagnostic cytology.
Potential of the in situ polymerase chain reaction in diagnostic cytology.
复制标题
原位聚合酶链反应在诊断细胞学中的潜力。
作者：
J. Pestaner;M. Bibbo;L. Bobroski;T. Seshamma;O. Bagasra
我们实验室通过光学显微镜对细胞学材料进行的常规检查包括特殊染色和酶组织化学，但一般不采用分子生物学技术。我们研究了利用原位聚合酶链反应（PCR）检查档案细胞学标本的可行性。为了采用经济且具有诊断相关性的技术，缩短了方案；提出请求后两天内就会得到结果。对移行细胞癌病例进行 p53 抑癌基因检查；直接掺入PCR的制备需要八个小时的实验室工作，热循环进行过夜，产物可视化需要第二天三个小时的实验室工作。用抗地高辛荧光和过氧化物酶探针在细胞质和核区域中发现扩增产物。原位 PCR 在诊断细胞学实验室中具有巨大的潜力。
Routine examination of cytologic material by light microscopy in our laboratory includes special stains and enzyme histochemistry, but molecular biology techniques are not generally employed. We examined the feasibility of utilizing the in situ polymerase chain reaction (PCR) for examination of archival cytologic specimens. Protocols were shortened in an effort to employ a technique that would be economical and have diagnostic relevance; a result would be obtained within two days of a request. Cases of transitional cell carcinoma were examined for the p53 tumor suppressor oncogene; preparation of direct incorporation PCR required eight hours of laboratory work, thermal cycling was performed overnight, and product visualization required three hours of laboratory work the following day. Amplification products were found in the cytoplasm and nuclear regions with an antidigoxigenin fluorescent and peroxidase probe. In situ PCR has enormous potential in the diagnostic cytology laboratory.