四种HBV DNA荧光定量PCR试剂比较及其结果分析

Assesement of three domestic commercial real-time polymerase chain reaction kits for detection of hepatitis B virus DNA using Qiagen Artus HBV TM PCR kit as reference

目的 评价3种国产HBV DNA荧光定量PCR试剂的特异性和灵敏度.方法 采用德国QIAGEN公司的Artus HBV DNA荧光定量PCR试剂(A)和国内生产的3种HBV DNA荧光定量PCR试剂(B、C、D)对203例HBV感染者血清标本进行平行检测,分析3种国产试剂与Artus试剂检测结果的差别及其相关性.结果 4种试剂检测203例血清标本HBV DNA的结果分别是A (5.78±1.78)、B(5.70±1.34)、C(5.51±1.96)、D(5.60±1.52),3种试剂与Artus试剂的相关系数分别为B:0.860、C:0.954、D:0.922. 对于HBV DNA载量高于106拷贝/ml的标本,3种国产试剂检测结果的均值与Artus无明显差异,但试剂B与Artus相关性明显不如试剂C和试剂D;对于HBV DNA载量在103～106 拷贝/ml的标本,试剂B和试剂C的均值水平与Artus试剂存在明显差别,试剂C和D与Artus试剂的相关性要优于试剂B;而对于30～103 拷贝/ml的标本,试剂B和试剂D的均值与Artus结果存在显著差异,且3种国产试剂与Artus试剂相关性很差.在26例30～103拷贝/ml样本中,3种国产试剂分别漏检8例(30.8%)、8例(30.8%)和11例(42.3%),7例<30拷贝/ml样本中3种国产试剂无一检出.结论 3种国产试剂的敏感性均较Artus试剂差,尤其是病毒载量低于103拷贝/ml的标本.在3种国产试剂中,试剂C与Artus试剂检测的结果相关性最好.

Objective To assess the specificity,sensitivity,accuracy of three domestic commercial real-time polymerase chain reaction kits for detection of hepatitis B virus DNA(HBV DNA real-time PCR kit)in comparison to Qiagen Artus HBV TM PCR kit as reference.Methods HBV DNA in 203 sera samples were detected by Artus HBV TM PCR kit of QIAGEN(A)and three domestic HBV DNA real-time PCR kit(B,C,D)in parallel,the sensitivity,specificity and accuracy of kits were determined by using the results of Qiagen Artus HBV TM PCR kit as reference.Results HBV DNA was detectable in 185 of 203 samplse by all four kits and the HBV DNA level measured by A,B,C,D are(5.78±1.78),(5.70±1.34),(5.51±1.96),(5.60±1.52)copies/ml,respectively.The correlation coefficient between HBV DNA level determined by Artus reagent(A)and the HBV DNA level of 185 samples measured by domestic kits B,C,D are 0.860,0.954,0.922,respectively.While the correlation coefficient remains high in samples with HBV DNA higher than 106 copies/ml,it drops significantly in samples with HBV DNA level lower and poor correlation was observed in samples with HBV DNA level lower than 103 copies/ml.Out of 26 samples with HBV DNA 30-103 copies/mL,the number of undetectable samples is 8(30.8%),8(30.8%)and 11(42.3%)by B,C,D kit,respectively.Among the 7 samples with lower level of than 30 copies/ml,all three domestic kits showed negative results.Conclusions The sensitivity and accuracy of three domestic kits is comparable with the Artus kit when samples with high HBV DNA level.However,all three kits need to be improved,especially to detect the HBV DNA lower than 103 copies/ml.