甲型H1N1流感超敏感诊断方法的建立及优化

Establishment and Optimization of Super-sensitive Diagnosis Method for Influenza A (H1N1)

利用几种助沉剂和核酸纯化方法富集病毒RNA,建立甲型H1N1流感超敏感诊断方法。收集31份甲型H1N1流感确诊病人提取的病毒RNA,稀释至1000分之一倍后分别应用酵母tRNA、糖原、AcrylCarrier助沉剂和磁珠、硅胶颗粒和离心柱的9种组合进行病毒RNA的富集,富集后分别进行RealtimePCR检测。并对检测结果进行统计学分析。利用几种助沉剂和核酸纯化方法的组合时,发现磁珠+AcrylCarrier助沉剂方法是最佳的核酸富集方法,能够大大提高病毒核酸的检出效率。建立了甲型H1N1流感超敏感诊断方法,该方法能够大大提高甲型H1N1流感的检出率。

To establish a super-sensitive diagnosis method for influenza A （ H1 N1 ） by enrichment of virus RNA using several settling agents and nucleic acid purification methods, virus RNA extracted from 31 confirmed influenza A （ H1 N1 ） patients were collected and diluted 1 000-fold. Then 9 combination of yeast rRNA, glycogen, Acryl Carrier settling agents and beads, silica particles and centrifugation column were used to enrich virus RNA. Then RNA were detected by Reahime PCR. It is resulted that Beads ＋ Acryl Carrier is found to be the best method of nucleic acid enrichment by combination of settling agents and nucleic acid purification method, which could greatly enhance the efficiency of viral nucleic acid detection.