人乳头瘤病毒分型液态芯片构建中影响因素-第三军医大学学报.DOC

人乳头瘤病毒基因分型液态芯片构建中的影响因素 党倩丽1，陈超2，杜篷2，杜艳华2，冯捷3 (1陕西省人民医院皮肤科, 陕西 西安 710068；2西北大学生命科学院, 国家微检测系统工程技术研究中心,陕西 西安 710069；3西安交通大学第二医院 陕西 西安710003) Influnecing factor of developing Human Papillomavirus types by using multiplexed suspension array DANG Qian-li1,CHEN Chao2, DU Pen2, Du Yan-hua2, Feng Jie3 1Department of Dermatology, Shannxi Provincial People’s Hospital ,Xi’an 710068, China, 2Northwest University, National Engineering Research Center for Miniturized Detection System,Xi’an 710069,China, 3Department of Dermatology,Second Hospital of Xi’an Jiaotong Universtiy, Xi’an, 710003 China 【Abstract】AIM: To investigate the Influnecing factor of developing genetypes for Human Papillomavirus by using multiplexed suspension array. MEDTHODS: Standard plasmids containing HPV genomic DNA developed a standard multiplexed suspension array for detection 13 HPV types of on LuminexTManalysis platform. the primer and probes were selected, HPV type-specific oligonucleotide probes were coupled to fluorescence-labeled polystyrene beads. Optimizing system of PCR; detecting the result of Coefficient of variation(CV) of standard HPV plasmids. RESULTS:Under a period of hybridation time, the hybridation time exert specific influence on result of hybridizing reaction. Concentration of PCR produce have little effect upon the Fluorescent intensity. CV range of HPV probes are 0.41%-14.69%. CONCLUSION: Hybridation time, pollution of sample in PCR, expiration date of microspheres, quality of probes have effect on the construction of detecting genetype of HPV by using multiplexed suspension array. 【摘要】目的：探讨在构建HPV基因分型液态芯片过程中的影响因素。方法：在LuminexTM平台上，采用国际公认的标准HPV质粒建立13种基因分型HPV 芯片，选择引物及探针，将探针有效的偶联到微球上。优化PCR体系，测定标准质粒变异系数(CV)值。结果：对构建芯片条件进行分析，杂交反应时间对杂交反应结果在一定的杂交时间内有明显的影响；PCR产物的浓度对荧光值没有明显影响；各型HPV探针CV范围在0.41%～14.69%之间。结论：杂交反应时间、样品PCR反应中的污染、微球的时效和探针的质量等均可以影响液态芯片系统的构建。 ------------------------------------- 收稿日期:2008 ;接受日期:2008- 基金项目：国家高技术研究发展计划（863）重大项目 （2002AA2Z2031）；陕西省自然科学基金项目(2000SM59) 作者简介：党倩丽 （1964-），女，陕西西安