促凝血与抗凝血对酶联免疫吸附实验中HBsAg检测结果的影响研究

Study on the Influence of Procoagulant and Anticoagulant on the Detection Results of HBsAg in Enzyme-Linked Immunosorbent assay

目的 分析促凝血和抗凝血样本对于酶联免疫吸附实验(ELISA)中乙肝病毒表面抗原(HBsAg)检测结果的影响。方法 选取2022年1月至2022年6月驻马店市中心血站共3 680例无偿献血者的血液样本进行研究,均留取血液样本两份,其中一份采取促凝血处理,另外一份采取抗凝血处理,所有血液样本使用的检测仪器及试剂均相同,并于相同条件下采取ELISA检测HBsAg;并依据HBsAg弱阳性参考品的比例加入抗凝剂开展测定;比较促凝血样本和抗凝血样本ELISA检测HBsAg的样本吸光度与标准吸光度比值(S/CO值)情况。结果 促凝血样本采取ELISA法检测HBsAg显示阳性48例,阳性率是1.30%;抗凝血样本采取ELISA法检测HBsAg显示阳性30例,阳性率是0.82%。促凝血样本用于ELISA法检测HBsAg的阳性率高于抗凝血样本,差异有统计学意义(P <0.05)。3 680例无偿献血者促凝血样本和抗凝血样本内共有18名的HBsAg检测结果存在不一致,促凝血样本HBsAg检测的S/CO值普遍高于抗凝血样本。经重复实验后,上述18例样本复试结果一致,均为阳性,重复实验结果和促凝血样本的检测结果相同。HBsAg质控血清依据比例加进抗凝剂后,伴随抗凝剂量不断升高,HBsAg含量以及S/CO值逐渐降低。促凝血样本用于ELISA法检测HBsAg的S/CO值高于抗凝血样本,差异有统计学意义(P <0.05)。结论 和促凝血样本相比,酶联免疫吸附实验中采取抗凝血样本检测HBsAg可能产生漏检的情况,建议留取促凝血样本开展检测,以确保检测结果的准确度。

Objective To analyze the effect of procoagulation and anticoagulation samples on the detection of hepatitis B virus surface antigen(HBsAg) in enzyme-linked immunosorbent assay(ELISA).Methods From January 2022 to June 2022,3680 unpaid blood donors in our central blood station were investigated.Two specimens were collected for each donor.One specimen was stored after the procoagulation treatment and one specimen after the anticoagulant treatment.The same testing instruments and reagents were used for all blood samples,and ELISA was used to detect HBsAg under the same conditions;and add anticoagulants according to the proportion of HBsAg weakly positive reference materials for testing.Comparison of sample absorbance to standard absorbance ratio(S/CO value) for ELISA detection of HBsAg between procoagulant and anticoagulant samples.Results ELISA was used to detect HBsAg in 48 cases of procoagulant samples,with a positive rate of 1.30%;30anticoagulant samples were tested for HBsAg using ELISA,with a positive rate of 0.82%.The positive rate of using procoagulant samples for ELISA detection of HBsAg is higher than that of anticoagulant samples,and the difference is statistically significant(P < 0.05).There were inconsistencies in the HBsAg test results of 18 out of 3680 voluntary blood donors,including both procoagulant and anticoagulant samples.The S/CO values of HBsAg test in procoagulant samples were generally higher than those in anticoagulant samples.After repeated experiments,the retest results of the 18 samples mentioned above were consistent and all were positive.The results of the repeated experiments were the same as those of the procoagulant samples.After adding anticoagulants to HBsAg quality control serum in proportion,the amount of anticoagulants continuously increases,and the HBsAg content and S/CO value gradually decrease.The S/CO values of coagulation promoting samples used for ELISA detection of HBsAg were higher than those of anticoagulant samples,and the difference was statistically significant(P < 0.05).Conclusion During the enzyme-linked immunosorbent assay,the missed diagnosis conditions of HBsAg could be found in the anticoagulant specimens.Thus the procoagulant specimen shall be kept to ensure the examination accuracy.