Poonnaniti A, Bhattarakosol P
Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
J Med Assoc Thai. 1996 Dec;79 Suppl 1:S96-103.
Infection with human papillomavirus (HPV) is an important etiological factor in the development of cervical cancer; detection of the viral genome is of prognostic importance, particularly for preneoplastic lesions. Nowadays, polymerase chain reaction (PCR) technique is a method of choice for HPV detection because of its sensitivity. However, providing a reliable diagnostic test, both sensitivity and specificity of the test should be evaluated. In the present study, purified plasmid HPV-16 DNA and HeLa-DNA, containing HPV-18 DNA, were amplified by PCR using L1 consensus primers specific for HPV. The amplified product was then analysed by gel electrophoresis (GE) and dot hybridization (DH). The generic oligonucleotide probes (GPs) labelled with Enhanced Chemiluminescence 3' labelling kit (ECL) were used in DH. The sensitivity of PCR reaction after determining by GE was 1 copy per cell for purified plasmid HPV-16 DNA and 20-80 copies per cell for HeLa-DNA while determining by DH was only 0.1 and 2-8 copies per cell, respectively. Thus, the detection of amplified product by DH using enhanced chemiluminescence system improved not only the specificity but also the sensitivity of HPV detection at least 10-fold.
人乳头瘤病毒（HPV）感染是宫颈癌发生发展的一个重要病因；病毒基因组的检测具有预后重要性，尤其是对于癌前病变。如今，聚合酶链反应（PCR）技术因其敏感性成为HPV检测的首选方法。然而，要提供一个可靠的诊断测试，该测试的敏感性和特异性都应进行评估。在本研究中，使用针对HPV的L1共有引物通过PCR扩增纯化的质粒HPV-16 DNA和含有HPV-18 DNA的HeLa-DNA。然后通过凝胶电泳（GE）和点杂交（DH）分析扩增产物。在点杂交中使用用增强化学发光3'标记试剂盒（ECL）标记的通用寡核苷酸探针（GPs）。通过GE测定时，纯化的质粒HPV-16 DNA的PCR反应敏感性为每个细胞1个拷贝，HeLa-DNA为每个细胞20 - 80个拷贝，而通过DH测定时分别仅为每个细胞0.1个拷贝和2 - 8个拷贝。因此，使用增强化学发光系统通过点杂交检测扩增产物不仅提高了特异性，还将HPV检测的敏感性至少提高了10倍。
