布鲁氏菌抗原检测试剂用国家参考品的研制

Preparation of national reference for detection reagent of Brucella antigen

目的研制布鲁氏菌抗原检测试剂评价用国家参考品。方法将3株布鲁氏菌标准株和10株阴性参考菌株在各自适宜的培养基和温度下进行培养,收集新鲜培养物并灭活,采用比浊法制备阳性和阴性参考品菌液;分光光度法制备最低检出量和重复性参考品菌液。对参考品进行均匀性检测和稳定性试验,组织4个实验室对布鲁氏菌抗原检测试剂用参考品进行协作标定。结果建立了由3株布鲁氏菌为阳性参考品、10株阴性菌为阴性参考品、最低检出量参考品及重复性参考品组成的布鲁氏菌抗原检测试剂评价用国家参考品。该参考品具有较好的均匀性及稳定性。协作标定结果为阳性参考品符合率均为100%,阴性参考品符合率均为90%,最低检出量参考品的检出限为1×10^7~1×10^6个/ml之间,重复性参考品均符合规定。结论研制的布鲁氏菌抗原检测试剂评价用国家参考品可用于布鲁氏菌抗原检测试剂的质量控制。

Objective To prepare national reference for detection reagent of Brucella antigen.Methods Three standard strains of Brucella and ten reference strains of non-Brucella were cultured in appropriate culture media at appropriate temperatures respectively.Fresh and uncontained cultures were collected,based on which positive and negative reference bacterial liquids were prepared by turbidimetry,and the reference bacterial liquids for minimum detection limit and repeatability were prepared by spectrophotometry.The prepared references were subjected to uniformity and stability test.Four laboratories were organized to coordinate the calibration of reference materials.Results Positive reference consisted of three samples of Brucella liquids,while negative reference consisted of ten samples of nonBrucella liquids.The reference for minimum detection and reproducibility consisted of frozen dried bacteria.The reference had good uniformity and stability.The collaborative calibration results of four laboratories showed that three positive references were 100%positive,ten negative references were 90%negative,and the lowest detection amount was between 10^7 cells/ml and 10^6 cells/ml.Repetitive reference test results were found to be consistent.Conclusion The reference can be used for quality control of Brucella antigen detection reagent.