自制汉坦病毒N蛋白IgM直接捕捉ELISA诊断试剂盒的初步评价  

Preliminary evaluation of self-made Hantavirus N protein IgM direct capture ELISA diagnostic kit

作　　者：姚苹苹[1] 朱函坪[1] 姚晨辉[2] 徐芳[1] 梅玲玲[1] 张政[1] 朱智勇[1] 邓小昭[3] 张云[3] 

机构地区：[1]浙江省疾病预防控制中心肾综合征出血热重点实验室,杭州310051 [2]大连医科大学 [3]南京军区军事医学研究所

出　　处：《中国媒介生物学及控制杂志》2010年第2期128-130,共3页Chinese Journal of Vector Biology and Control

基　　金：浙江省医药卫生科学研究基金(2006B020);国家"863"高技术研究发展计划(2007AA02Z465)~~

摘　　要：目的初步评价应用自制辣根过氧化物酶(HRP)标记汉坦病毒(HV)重组N蛋白(rNP)的rNP-IgM直接捕捉ELISA诊断试剂盒。方法考核该试剂盒的特异性和稳定性;比较其与同类商品试剂盒的临床检测结果,以评估检测血清抗HV-IgM的敏感性。结果(1)该试剂盒仅与抗HV-IgM阳性血清发生反应,而与抗水痘病毒-IgM(抗VZV-IgM)、抗流行性乙型脑炎病毒-IgM(抗JEV-IgM)、抗登革热病毒-IgM(抗DV-IgM)、抗手足口EV71病毒-IgM(抗EV71-IgM)阳性血清均无反应;试剂盒放置37℃3d,检测血清抗HV-IgM的活性,无明显降低。(2)在120例临床疑似肾综合征出血热的144份血清中,2种试剂盒仅对12例的12份血清抗HV-IgM检测结果不一致,其中8例的首份血清,该试剂盒阳性,商品试剂盒阴性(第2份血清2种试剂盒均阳性);3例的首份血清(未采到第2份血清),该试剂盒为临界值,商品试剂盒阴性;另1例的首份血清该试剂盒阳性,商品试剂盒阴性(2种试剂盒第2份血清和第3份血清均阳性)。结论实验室研制的捕获法ELISA抗HV-IgM诊断试剂盒具有良好的特异性、稳定性和敏感性,适于HV感染的临床诊断和流行病学监测。Objective To provide a preliminary evaluation of the self-made horseradish peroxidase （HRP） marked Hantavirus （HV） recombinant N protein （rNP） rNP- IgM direct capture ELISA diagnostic kit. Methods Assessment of the specificity and stability of the kit and comparison of clinical results with similar products were performed to evaluate the sensitivity of the kit in the detection of serum anti-HV-IgM. Results （1） The kit was only responsive to anti-HV-IgM positive serum, and irrcsponsive to anti-varicella-zoster virus-IgM （anti-VZV-IgM）, anti-Japanese encephalitis virus-IgM （anti-JEV-IgM）, anti-dengue virus-IgM （anti-DV-IgM）, anti-hand, foot and mouth EV71 virus-IgM （anti-EV71-IgM） positive sera. No obvious reduction in serum anti-HV-IgM detecting capability was noticed after placement at 37 ℃ for 3 d. （2） In 144 sera samples in 120 patients with suspected hemorrhagic fever with renal syndrome, inconsistency was observed only in the anti-HV-IgM test results in 12 sera of 12 patients between the two kinds of kits, in which 8 primary sera samples were detected positive by the kit and negative by commercial ones （the secondary sera samples were positive for both kits）; 3 primary samples （the secondary samples unavailable） were at the critical value for the kit and negative for commercial kits. Meanwhile, one other primary serum sample was positive for the kit and negative for the commercial ones （the secondary and tertiary ones positive for both）. Conclusion The laboratory developed capture ELISA anti-HV-IgM diagnostic kits had favorable specificity, stability and sensitivity, suitable for the clinical diagnosis and epidemiological surveillance of HV infections.

关 键 词：汉坦病毒 N蛋白 重组表达 IgM捕捉ELISA 

分 类 号：R373.32[医药卫生—病原生物学]