低密度基因芯片技术检测人乳头瘤病毒基因型的方法研究

目的:构建并评价检测人乳头瘤病毒(HPV)基因型的低密度基因芯片方法。方法:将15种高危、5种低危和Cp8304共21种HPV型特异寡核苷酸探针加尾、固定在尼龙膜上,制成低密度基因芯片。以第二代杂交捕获(HC2)法作为检出HPV的金标准,随机选取148例标本,用低密度基因芯片技术进行HPV检出和基因分型,并对检测系统的敏感性、特异性及重复性进行测试,判定基因测序检测芯片的可靠性。结果:148例标本,采用低密度基因芯片与HC2检出的完全符合率是95.95%(Kappa值为0.82),共检出18种HPV基因型。灵敏度、特异性较好,检测系统稳定。结论:低密度基因芯片技术具有敏感、特异、快速、简便、经济等特点,一次检测既能测定HPV感染又能进行HPV基因分型,对临床HPV的筛查、诊治和预后评估具有较大价值。

Identification of human papillomavirus genotypes by low - density gene chip detecting method

Objective： To develop a low-density gene chip method for detection of human papillomavirus （HPV） genotypes. Methods： Twenty-one HPV type-specific oligonucleotide probes, which containing 15 subtypes of high-risk HPV, 5 subtypes of low-risk HPV types and the subtype of CP8304, were tailed and immobilized on a nylon membrane to develop the low-density gene chip-based detecting system. Hybrid Capture 2 （HC2） was used as a gold standard to evaluate the new detecting system. A total of 148 cervical samples were selected by HC2 and were further detected with the low-density gene chip method. The sensitivity, specificity and repeatability of the detectiing system were evaluated, and the reliability of genotypes was tested by DNA sequencing. Results： The low-density gene chip detecting result showed a good agreement with that of HC2. The observed agreement was 95.95% with kappa = 0.82. A total of 18 genotypes were detected. Good sensitivity and specificity as well as stabilized system were found. Conclusion： The Low-density gene chip detection appears to be a high specific, sensitive, rapid, simple and economic approach used for HPV screening, diagnosis and prognosis evaluation.