Analysis of suspicious results of serum HBV DNA detected by fluorescence quantitative PCR
10.3760/cma.j.issn.1009-9158.2013.03.006
- VernacularTitle:实时荧光定量PCR检测血HBV DNA的疑难结果分析及其对策
- Author:
Zhanguo CHEN
;
Wu ZHOU
;
Zhongyong WANG
;
Yalei JIN
;
Zhihua TAO
- Publication Type:Journal Article
- Keywords:
Polymerase chain reaction;
Hepatitis B virus;
DNA,viral
- From:
Chinese Journal of Laboratory Medicine
2013;(3):217-221
- CountryChina
- Language:Chinese
-
Abstract:
Objective To analyze the suspicious results of serum HBV DNA by fluorescence quantitative PCR and develop appropriate countermeasures in order to improve the quality of detection of HBV DNA.Methods Blood samples of patients from the First Affiliated Hospital of Wenzhou Medical College from 2008 to 2011 were analyzed for HBV DNA by fluorescence quantitative PCR.1969 cases of suspicious results,judged by the rule of review the results of serum HBV DNA combined with the historical results,PCR amplification curve,HBV serum markers and clinical diagnosis,were analyzed and redetected by using of two different reagents,careHBV PCR Kit and careHBV PCR Kit V2,at the same time.The consistency and inconsistency ratio of the results were evaluated.Both the reasons of inconsistent and the undetected rates of careHBV PCR Kit were analyzed.The two reasons for the inconsistent results included the reagent related factors,e.g,showing no amplification curve caused by the false negative and abnormal low efficiency of amplification curve,and the non reagent related factors such as operating pollution and other sample factors.Results There were 115 154 blood samples were detected for HBV from 2008 to 2011 and 1969 samples (1.71%) with suspicious results were redetected.The consistency and inconsistency results were 1588 (80.65%) and 381 (19.35%),respectively.Every year from 2008 to 2011,the percentage of the inconsistent results caused by the reagent related factors were 18.87%,20.23%,51.33% and 59.57% respectively,which showed an increasing trend,and the percentage of inconsistent results caused by the nonreagent related factors were 81.13%,79.77%,48.67% and 40.43% respectively,which showed a declining trend year by year.The undetected rates of careHBV PCR Kit were 2.49%,4.08%,10.09% and 14.47% respectively,showing an increasing trend.Conclusions The redetection for the specimens with the suspicious results by using of different reagents can avoid the blind detection of HBV DNA and reduce the experimental error.All the clinical samples for quantitative HBV DNA including the mutations of HBV gene can be measured accurately and effectively,which is helpful to hepatitis B patients for antiviral therapy.