不同ELISA系统检测人血清百日咳IgG抗体的比较研究

A comparative study of different ELISA systems for detection of IgG antibodies against pertussis in human serum

目的使用A、B、C 3组酶联免疫吸附试验(ELISA)系统进行百日咳疫苗临床血清检测,每组ELISA系统包括至少一种百日咳毒素(PT)、一种丝状血凝素(FHA)和一种黏附素(PRN)包被抗原,其中C组抗原包括2种FHA抗原,酶标二抗是相同的抗人IgG抗体。比较不同包被抗原对百日咳疫苗临床血清中3种IgG抗体检测结果的影响。方法收集164对无细胞百日咳、白喉和破伤风疫苗基础免疫前、免疫后临床血清,使用ELISA法检测和计算血清百日咳IgG抗体的转阳率和单位值。使用χ2检验比较转阳率差异,使用方差分析比较抗体检测结果。结果A组抗原检测IgG抗体结果为PT 82.19IU/mL,FHA 81.66IU/mL,PRN39.34IU/mL;转阳率均为100.0%。B组抗原检测结果为PT 95.59IU/mL,FHA 57.39IU/mL,PRN 40.76IU/mL;转阳率PT抗体为99.4%,其余为100.0%。C组抗原检测结果为PT 60.74IU/mL,2种FHA分别为29.33IU/mL和53.32IU/mL,PRN 34.05IU/mL,转阳率为99.4%、95.1%、100.0%和98.8%。3组抗原检测的临床血清转阳率经χ2检验FHA差异有统计学意义(P<0.05),PT和PRN差异无统计学意义(P>0.05);抗体GMC经方差分析,所有组抗原检测3个抗体值之间差异均有统计学意义(P<0.05)。结论本次人临床血清抗体转阳率和抗体值达到保护力水平,但是不同抗原检测系统之间差异均有统计学意义,应尽快建立百日咳抗血清IgG检测用抗原试剂国家参考品。

Objective The clinical serum of pertussis vaccine was detected by using group A,B and C ELISA systems.Each group of ELISA systems consisted of at least one pertussis toxin(PT),one filamentous hemagglutinin(FHA)and one adhesin(PRN)coated antigen.Group C antigens included two FHA antigens,and the second enzyme labeled antibody was the same anti-human IgG antibody.The purpose of this study was to compare the effects of different coated antigens on the detection of three IgG antibodies in clinical serum of pertussis vaccine.Methods Totally 164 pairs of clinical sera of acellular pertussis,diphtheria and tetanus vaccines were collected before and after basic immunization.The positive conversion rate and unit value of serum pertussis IgG antibody were detected and calculated by ELISA.The difference of positive conversion rate was compared byχ2 test,and the results of antibody detection were compared by variance analysis.Results The results of IgG antibody detection by group A antigen were PT 82.19 IU/mL,FHA 81.66 IU/mL,PRN 39.34 IU/mL,and the positive conversion rates were 100.0%.The results of antigen detection in group B were PT95.59 IU/mL,FHA 57.39 IU/mL,PRN 40.76 IU/mL.The positive conversion rate of PT antibody was 99.4%,and the others were 100.0%.The results of antigen detection in group C were PT 60.74 IU/mL,FHA 29.33 IU/mL and 53.32 IU/mL,PRN 34.05 IU/mL,respectively.The positive conversion rates were 99.4%,95.1%,100.0%and 98.8%.There was significant difference in FHA among the three groups byχ2 test(P<0.05),there was no significant differencein PT and PRN among the three groups(P>0.05),and there was significant difference among the three antibody values of all groups by variance analysis(P<0.05).Conclusion The positive conversion rate and antibody value of human clinical serum reached the level of protection,but there were significant differences among different antigen detection systems.The national reference for antigen reagent for pertussis antiserum IgG detection should be established as soon as possible.