百日咳抗体酶联检测试剂盒的研制及其应用

为了研制百日咳抗体酶联检测试剂盒以调查吉林省2~8岁儿童百日咳、白喉及破伤风抗体水平。采用百日咳菌液经硫酸铵盐析、PBS溶液浸提及蔗糖密度梯度离心提取的PT和FHA作为包被抗原,辣根过氧化物酶(HRP)标记羊抗人IgG,作为抗体制备ELISA试剂盒。并经敏感性、特异性、重复性试验考查该试剂盒质量。结果显示,试剂盒敏感性可达0.0036 IU/m l;重复性较好,CV<4%。试剂盒置于37℃3d敏感性无明显变化。吉林省2~8岁儿童白喉、破伤风的抗体水平较高,百日咳的抗体水平相对较低。此方法制备的百日咳抗体酶联检测试剂盒的质量符合要求,可应用于临床检验与流行病学监测。

Preparation and application of ELISA kit in detection of human antibodies against pertussis

The ELISA kit was prepared by using PT and FHA as coating antigens,which were purified by salting out with ammonium sulfate from pertussis culture supernatant,soaking by PBS solution and sucrose density gradient centrifugation.HRP-labled sheep anti-human IgG as signal antibody.The sensitivity,specificity and reproducibility of the kit were studied.The kit had good sensitivity(0.0036 IU/ml),specificity and reproducibility(CV<4%).After the kit stored at 37℃ for 3 days,no significant change was observed in relation to it's sensitivity.On the basis of the developed ELISA kit,the specific antibody for pertusis is detectable particularly in presence of high levels antibodies responses to diphtheria and tetanus among 2 to 8 year old children in Jilin provnice.