蛋白免疫印迹实验MP试剂快速法与隔夜孵育法的比对结果分析

Comparison on rapid test of MP reagent of Western blot and overnight incubation method

目的为了减少艾滋病病毒(HIV)确证实验室"不确定"样本的例数,使感染早期病人得到及时诊断,晚期病人得到及时治疗。方法对2012-2013年HIV-1抗体筛查阳性血清样本,用蛋白免疫印迹试验(WB)MP试剂快速法进行确证检测,确证结果为"不确定"的样本,采用MP试剂隔夜孵育法与之进行了比对。结果 46例WB"阳性-不确定"样本中,22例"不确定-阴性"及4例"不确定-失访"样本,其隔夜孵育法与快速法相比,确证条带数均未增加,部分样本颜色有不同程度的加深。20例"不确定-阳性"样本,有10例条带数在原有的基础上有不同程度的增加,并完全满足国家阳性判断标准,无需随访就可以确证为"阳性"结果;另10例条带在原有的基础上均有加深,其中3例符合阳性判断标准。对于20例"不确定-阳性"样本,隔夜孵育法与快速法差异有显著的统计学意义(P<0.05)。结论隔夜孵育法与快速法相比,反应时间较长,血清中的抗体与纤维膜上的抗原结合比较充分,一些早期或晚期感染病例在快速法中不显现的条带在隔夜孵育法得到了充分的展现。建议确证实验室如采用MP试剂快速法的,当结果为"阳性-不确定"的情况下,同时筛查结果为三种以上阳性者,应采用隔夜孵育法进行复测确证实验。

Objective To reduce the number of undefined samples from laboratory, for the early diagnosis of in- fected patients and timely treatment of patients in the terminal stage. Methods Western blot（WB） MP reagent rapid method was used to confirm the ＂indeterminate＂ samples among serum samples from human immunodeficiency virus type 1 （HIV-1） antibody-positive individuals during 2012-2013, compared to MP reagent overnight incubation method. Results Among forty six cases of ＂positive but indeterminate＂ samples, including 22 negative cases and 4 cases ＂lost to follow-up＇,overnight incubation method compared with the rapid method confirmed bands were all not increased, only color of parts of samples had different degrees of deepening; 20＂ uncertainty positive＂ samples, bands of 10 cases were increased at some degree, and conformed to the national standard of positive judgment, con- firmed as ＂positive＂ results without follow-up; bands of the other 10 cases were deepened, of which three cases con- formed to positive judgment standard; for 20 cases of＂uncertainty positive＂ samples, there was significant difference between overnight incubation method and fast test method（P〈0.05）, there was statistical significance. Conclusion Compared to fast rest method, overnight incubation method had more benefits, such as long response time, and binding of antibody in serum and the fiber membrane antigen was full, some strips of early or late infections with no appearance in the rapid method was fully displayed in the overnight incubation method; suggested that samples of HIV-1 antibody screening from laboratory were confirmed by MP reagent method, when the ＂positive - not sure＂ situations, such as screening results at the same time as more than three kinds of positive, recommended the use of overnight method for retesting the confirmatory tests.