论文部分内容阅读
目的建立一种新的可以快速筛查急性肝炎患者HBV和HCV感染的多重实时荧光定量PCR方法。方法收集596例急性肝炎患者血清或血浆样本,应用自主构建的多重实时荧光定量PCR方法同时检测HBV和HCV,同时应用HBV和HCV单管检测试剂进行检测分析。结果新建立的多重实时荧光定量PCR体系,可以有效地检测HBV和HCV的阳性质控品及阳性样本。多重实时荧光定量PCR体系特异性较好,检测HBV和HCV时,两者之间无交叉反应,且以多种血浆病毒的核酸样品为模板进行检测时结果全为阴性;多重实时荧光定量PCR体系检测HBV、HCV的灵敏度分别为20 IU/ml、50 IU/ml;多重实时荧光定量PCR体系精密度好,检测HBV、HCV的CV值分别为0.11%、0.096%。在检测临床样本时,多重复合体系检测血清或血浆中HBV和HCV的检出率与单独检测HBV和HCV具有很好的一致性,且HBV和HCV的一致率分别达到97.7%(k=0.952)、98.8%(k=0.976);同时有6例样本出现HBV和HCV共感染。结论新型多重实时荧光定量PCR方法对快速检测及筛查HBV和HCV较快捷,而且特异性好、灵敏度高、精密度好。因此多重实时荧光定量PCR方法对2种肝炎病毒的检测具有重要意义。
Objective To establish a new multiplex real-time fluorescence quantitative PCR method for rapid screening of HBV and HCV infection in patients with acute hepatitis. Methods Serum or plasma samples from 596 patients with acute hepatitis were collected. HBV and HCV were detected by multiplex real-time PCR. The HBV and HCV single-tube detection reagents were also tested. Results The newly established multiplex real-time PCR system could effectively detect positive control samples and positive samples of HBV and HCV. The multiplex real-time PCR system has better specificity. When HBV and HCV were detected, there was no cross-reaction between the two and the results were negative when using a variety of samples of plasma virus nucleic acid as a template. Multiple real-time fluorescence quantitative PCR The sensitivity of detection of HBV and HCV were 20 IU / ml and 50 IU / ml, respectively. The multiplex real-time PCR system had good precision and the CV values of detecting HBV and HCV were 0.11% and 0.096% respectively. The detection rate of HBV and HCV in serum or plasma by multiplex system was very consistent with that of HBV and HCV alone when detecting clinical samples. The concordance rates of HBV and HCV were 97.7% (k = 0.952) , 98.8% (k = 0.976). In the meantime, 6 cases were co-infected with HBV and HCV. Conclusion The new multiplex real-time PCR method is rapid and sensitive for the rapid detection and screening of HBV and HCV with good specificity, high sensitivity and good precision. Therefore, multiple real-time fluorescence quantitative PCR detection of two kinds of hepatitis virus is of great significance.