基因芯片检测输血传播病毒方法的实验研究

目的建立基因芯片快速检测经输血传播病毒核酸的方法,进而探讨该方法用于检测临床标本的可行性。方法通过PCR获得TTV病毒ORF1基因的DNA片段,克隆,从重组质粒扩增DNA片段,并点到玻璃载体上,制成芯片。与TTV病毒、甲型肝炎病毒、乙型肝炎病毒及戊型肝炎等病毒的PCR产物进行杂交,以检测探针的特异性。结果该基因芯片探针仅与TTV毒株的PCR产物杂交呈阳性,与对照病毒的PCR产物杂交呈阴性。敏感性试验显示,用该方法检测了27份疑似TTV临床病料,22份阳性;而用PCR法扩增TTV ORF1基因确诊为阳性的只有19份。结论利用基因芯片检测TTV的PCR产物,特异性和敏感性强,可作为TTV临床标本检测方法。

Detection of transfusion transmitted virus gene chip method of experimental study

Objective To establish the gene chip method of rapid detection of TTV DNA, and then explore the method used to detect the feasibility of clinical samples. Methods TTV virus ORF1 gene PCR to obtain the DNA fragments, cloning, DNA fragments amplified from the recombinant plasmid,and point to the glass carrier,made of silicon. And Trv virus, hepatitis A virus, hepatitis B virus and hepatitis E vires like PCR products were hybridized to detect the probe specificity. Results The results of the gene-chip probe only with Trv strains positive for PCR product of hybridization with the control virus and organizations PCR product of hybridization were negative. Sensitivity tests showed that Using this method to detect clinical disease, 27 were suspected of TTV material, 22 were positive ; but with the TTV ORF1 gene was amplified by PCR confimed positive, only 19 copies. Conclusion The detection of TTV DNA microarray of the PCR product specificity and sensitivity of strong, repeatable, can be used as clinical samples TTV detection methods.