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目的 建立免疫血清中抗b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)磷酸多聚核糖基核糖醇(Polyri-bosylribitol phosphate,PRP)抗体体外血清杀菌试验(Serum bacterialcid assay,SBA)方法。方法通过对补体和指示菌株的筛选,建立体外测定血清中抗Hib-PRP抗体杀菌试验方法,并对实验体系的耐变性、特异性及精密性进行验证。结果指示菌Hib-EAGAN株与Hib阳性血清显示出良好的杀菌特异性,其平均非特异性杀菌率为0;Pel-Freez公司提供的4批补体中,批号为17830的补体具有良好的杀菌稳定性和特异性;经验证,建立的Hib-SBA体系具有良好的耐变性和特异性,其精密性CV值在16%～28%之间。结论成功建立了血清抗Hib-PRP抗体杀菌活性体外检测方法,该方法具有检测样本量大、省时、易标准化等优点。
OBJECTIVE: To establish a serum in vitro seropreservative assay (SBA) method against Haemophilus influenzae type b (Hib) polyribosylribitol phosphate (PRP) in immune serum. Methods The anti-Hib-PRP antibodies in serum were tested by the screening of complement and indicator strains. The bactericidal test of anti-Hib-PRP antibody in serum was established. The anti-degeneration, specificity and precision of the system were also verified. Results The indicator bacteria Hib-EAGAN and Hib-positive sera showed good bactericidal specificity with an average nonspecific bactericidal rate of 0; of the 4 lots provided by Pel-Freez, the complement of batch 17830 had good bactericidal stability And specificity. The established Hib-SBA system has good resistance to variability and specificity, and the CV value of the precision is between 16% and 28%. Conclusion The in vitro detection method of bactericidal activity of anti-Hib-PRP antibody was successfully established. The method has the advantages of large sample size, time-saving and easy standardization.