摘要
目的建立一种运用PCR结合基因芯片技术检测和鉴定引起腹泻的常见肠道致病菌方法。方法根据生物信息学技术以细菌16S rDNA和23S rDNA序列设计各细菌的特异型探针和引物,在基因芯片制备基础上,通过对靶基因的扩增、杂交和对杂交结果的分析,对常见肠道致病菌的检测和鉴定效果进行评价,同时对临床采集100份腹泻患者粪便标本通过细菌培养结果,比较基因芯片技术的灵敏性和特异性。结果 100份临床粪便标本中,采用基因芯片技术共检出50份(50%)携带肠道致病菌;细菌培养鉴定出47份(47%)携带肠道致病菌,其中42份基因芯片与细菌培养结果一致,准确率为79.25%,与基因芯片比较差异无统计学意义(χ2=5.28,P>0.05)。结论我们所建立的基因芯片系统具有高通量和高准确性,可以作为临床鉴定细菌的一个重要的工具,而且适用于流行病学调查研究。
Objective To establish a PCR-Gene chip technology for detecting common intestinal pathogens that causes the diarrhea and provide the theory instruction to the intestinal tract inflammation. Methods Biotin-labeled universal primers for amplification of bacterial 16S rDNA and 23S rDNA were used to amplify extracted genome DNA. Appraising the detect and authenticate effect through expanding the target gene, the hybrid and the hybrid result analysis, meanwhile 100 fecal samples from patients with diarrhea were detected and identified using this assay and bacilliculture results, to inspect the sensitivity and specificity of gene chip technology. Results Among 100 fecal samples, 50 （50%） samples were posi- tive for the intestinal pathogens by the PCR-Gene chip method; nevertheless 47 （47%） samples were positive for the intes- tinal pathogens by bacterial culture ; and the results of gene chip method were consistent with bacterial culture in 42 samples and the coincidence rate was 79.25%. Conclusions The gene chip system we established has high flux and the high ac- curacy, which is a rapid and accurate method to detect the common intestinal pathogens, and it is also well suitable for the epidemiology investigation and study.
出处
《山东医药》
CAS
2013年第4期24-26,29,I0001,共5页
Shandong Medical Journal
