幽门螺杆菌尿素酶B亚单位特异性抗体检测方法的建立与评价

目的建立并评价幽门螺杆菌(Hp)尿素酶B亚单位(UreB)特异性抗体的检测方法。方法应用纯化的重组HpUreB作为包被抗原,建立检测HpUreB特异性抗体的ELISA间接法。以Hp抗体Western blot检测作为“金标准”,评价所建立方法的真实性和可靠性。结果检测血清特异性IgG的灵敏度为100·0%,特异度为97·7%,阳性预测值为97·2%,阴性预测值为100·0%,约登指数为0·977,符合率为98·7%,试验的一致率为98·5%;检测血清中总的特异性Ig灵敏度为97·1%,特异度为95·3%,阳性预测值为94·4%,阴性预测值为97·6%,约登指数为0·925,符合率为96·2%,试验的一致率为97·8%;检测唾液中特异性sIgA的灵敏度为96·2%,特异度为94·5%,阳性预测值为89·3%,阴性预测值为98·1%,约登指数为0·907,符合率为95·1%,试验的一致率为97·5%;检测粪便标本中的特异性sIgA的灵敏度为92·0%,特异度为90·2%,阳性预测值为85·2%,阴性预测值为94·9%,约登指数为0·822,符合率为90·9%,试验的一致率为98·6%,CV值均小于15%。结论该检测方法真实性、可靠性、重复性良好,能满足临床标本检测的要求。

Development and Evaluation of Method for Determination of Specific Antibody against Helicobacter pylori Urease Subunit B

Objective To develop and evaluate the method for determination of specific antibody against Helicobacter pylori(Hp) Urease subunit B(UreB).Methods Develop an indirect ELISA method for determination of specific antibody against Hp UreB using purified recombinant Hp UreB as coating antigen.Evaluate the reality and reliability of developed mthod using Western blotting of Hp antibody as golden standard.Results The sensitivity,specificity,positive predictive value,negative predictive value,Youden index,accuracy and consistency of the developed method were 100%,97.7%,97.2%,100%,0.977,98.7%,98.5% for determination of serum specific IgG,97.1%,95.3%,94.4%,97.6%,0.925,96.2% and 97.8% for serum specific Ig,96.2%,94.5%,89.3%,98.1%,0.907,95.1% and 97.5% for sIgA in saliva,and 92.0%,90.2%,85.2%,94.9%,0.822,90.9% and 98.6% for sIgA in feces,respectively.Conclusion The developed indirect ELISA method showed good reality,reliability and reproducibility and met the requirements for determination of specimens in clinic.