溶血、脂血和黄疸对磁珠法乙型肝炎病毒核酸检测试剂检测下限的影响

Lower detection limit of novel HBV NAT kits in hemolysis,lipemia and astragalus blood:magnetic bead method

目的评估不同血红蛋白(Hb)、甘油三酯(TG)、总胆红素(TBil)浓度的血浆标本,对最新一代国产磁珠法HBV DNA检测试剂检测下限的影响。方法挑选临床不同血浆性状的HBV阴性血液标本,按照一定比例加入HBV国家标准品,使标本中的HBV DNA终浓度达到检测试剂最低检测限。再对标本中Hb、TG以及TBil进行定量检测,根据检测浓度将标本进行分组,性状正常的标本作对照。之后进行HBV DNA核酸检测,计算各组HBV DNA检出率,使用单因素方差分析进行组间差异比较。结果血浆游离Hb浓度在10.13±1.07g/L水平时,HBV DNA不能被完全检出,高TG和高TBil组HBV DNA检出率未受影响,Ct值差异无统计学意义(P<0.05)。结论在使用最新一代国产磁珠法HBV DNA检测试剂时,脂血、黄疸以及轻中度溶血标本均可正常进行检测,但当Hb达到临床重度溶血标准时,检测结果存在假阴性可能,应重新抽取血液标本。

Objective To evaluate the effects of different hemoglobin(Hb),triglyceride(TG)and totalbilirubin(TBil)on the lower detection limits of the latest generation of domestic HBV DNA detection kits using magnetic bead method.Methods HBV-negative blood samples with different clinical plasma traits were selected and spiked with HBV national standard products according to a certain ratio,so that the final concentration of HBV DNA in the sample reached the minimum detection limit of the detection kits.The samples,after quantitatively tested for Hb,TG and TBil,were grouped according to the detection concentration,and the samples with normal traits were enrolled as controls.Afterwards,HBV DNA nucleic acid detection(NAT)was performed,and the NAT-yield rate in each group was calculated and compared using one-way analysis of variance(ANOVA)Results HBV DNA could not be completely detected when plasma free hemoglobin concentration was(10.13±1.07)g/L,and HBV NAT-yield rate in high TG and TBil group was not affected.There was no significant difference in Ct value.(P<0.05).Conclusion The latest generation of domestic HBV DNA detection kits could be applied to lipemia,jaundice and mild to moderate hemolysis blood samples,whereas false negative results may occur in severe hemolysis samples,and blood sample should be re-drawn for retest in such condition.