Sayan Murat, Willke Ayşe, Meriç Meliha, Etiler Nilay
Kocaeli Universitesi Tip Fakültesi Hastanesi, Merkez Laboratuvari, PCR Unitesi, Kocaeli.
Mikrobiyol Bul. 2009 Jan;43(1):103-14.
External quality assessment (EQA) has been playing an increasingly important role in the implementation of nucleic acid amplification techniques (PCR) for clinical diagnosis. In this study, the results of HBV-DNA quantification and HCV-RNA detection tests evaluated by United Kingdom National External Quality Assessment Scheme for Microbiology (NEQAS) were analysed and the performance of our laboratory was evaluated. Between April 2006-January 2008, in four different distribution panels including 16 freeze-dried serum and six plasma specimens for HBV-DNA and HCV-RNA testing, respectively, were received. Viral nucleic acids were extracted by magnetic particle technology (NucliSENS-easyMAG, bioMérieux, Boxtel, Holland). HBV-DNA and HCV-RNA tests were performed by Fluorion HBV quantitative v2.0 and Fluorion HCV quantitative v2.1 (lontek AS, Istanbul, Turkey) kits in real-time PCR (iCycler IQ v3.0a - BioRad Laboratories, Hercules, CA, USA) platform. The performance scores of all the quantification tests of HBV-DNA were 2 (completely correct result) and a strong correlation (r= 0.987) between the quantitative HBV data and the target values was found by linear regression analysis. The NEQAS scores of HCV-RNA testing, except for a false negative result (since the viral load in this specimen was very low--79 IU/ml--it was not scored by NEQAS), were 2 in all specimens. The evaluation of the data revealed 100% detection in HBV-DNA and 83.3% detection in HCV-RNA. In conclusion, the results of this study showed high accuracy of HBV quantification in the samples of HBV infected patients under antiviral therapy. However, the analytical sensitivity of HCV-RNA quantitative kit should be improved for the purpose of reliable HCV-RNA results. External quality control panels are important tools for monitoring the quality of diagnostic laboratory tests. Therefore, PCR laboratories should always have EQA in routine procedures.
外部质量评估(EQA)在临床诊断中核酸扩增技术(PCR)的应用中发挥着越来越重要的作用。在本研究中,分析了由英国国家微生物学外部质量评估计划(NEQAS)评估的HBV-DNA定量和HCV-RNA检测试验结果,并评估了我们实验室的性能。在2006年4月至2008年1月期间,分别收到了四个不同的分发小组的样本,其中包括16份用于HBV-DNA检测的冻干血清样本和6份用于HCV-RNA检测的血浆样本。采用磁珠技术(NucliSENS-easyMAG,生物梅里埃公司,荷兰博克斯泰尔)提取病毒核酸。在实时PCR(iCycler IQ v3.0a - 伯乐生命医学产品公司,美国加利福尼亚州赫拉克勒斯)平台上,使用Fluorion HBV定量v2.0和Fluorion HCV定量v2.1(土耳其伊斯坦布尔的Lontek AS公司)试剂盒进行HBV-DNA和HCV-RNA检测。所有HBV-DNA定量检测的性能得分均为2(结果完全正确),通过线性回归分析发现定量HBV数据与目标值之间存在强相关性(r = 0.987)。除了一个假阴性结果(由于该样本中的病毒载量非常低 - 79 IU/ml - NEQAS未对其进行评分)外,所有样本的HCV-RNA检测的NEQAS得分均为2。数据评估显示,HBV-DNA检测率为100%,HCV-RNA检测率为83.3%。总之,本研究结果表明,在接受抗病毒治疗的HBV感染患者样本中,HBV定量具有很高的准确性。然而,为了获得可靠的HCV-RNA结果,应提高HCV-RNA定量试剂盒的分析灵敏度。外部质量控制样本是监测诊断实验室检测质量的重要工具。因此,PCR实验室在常规程序中应始终进行EQA。