【摘要】 目的探讨三磷酸腺苷(Adenosine triphosphate,ATP)生物发光快速微生物检测法应用于疫苗中间品无菌试验的可行性。方法测定各种培养基与疫苗培养上清液(不含细胞碎片)的本底值,并确定阳性限值;确定测试仪的检测限及线性范围;考察非细菌来源的ATP对测定值的影响;筛选适宜培养基;比较ATP生物发光法与无菌试验检测结果的一致性。结果除199培养基外,其他培养基的RLU本底值均低于205,阳性限值为500;测试仪的最低检测限为10-15mol/L,在10-15～10-6mol/L范围内,线性关系良好;采用增菌法进行试验,可不考虑非细菌来源的ATP的影响;筛选出的适宜培养基为硫乙醇酸盐流体培养基(不含琼脂);ATP生物发光法和无菌试验检测结果的阳性符合率为80%,2种方法各有1个样品另一种方法未检出。结论 ATP生物发光法操作简便,应用范围广泛,可应用于疫苗中间品的无菌试验。更多还原

【Abstract】 Objective To investigate the feasibility of rapid determination of microorganisms by ATP bioluminescence assay used for sterility test of intermediate product of vaccine. Methods The background RLUs of various media and vaccines were determined, based on which the positive cutoff value was obtained. The detection limit and linear range of detector were determined, and the effect of non-bacterium-derived ATP on determination result was investigated. The optimal medium was screened, and the consistency of results of ATP bioluminescence assay and sterility test was evaluated. Results All the background RLUs of media except 199 medium were less than 205, and the positive cutoff value was 500. The detection limit and linear range of detector were 10-15 mol / L and 10-15 ～ 10-6 mol / L respectively. The effect of non-bacterium-derived ATP on determination result was inconsiderable when bacterial enrichment test was used. The agar-free thioglycollate fluid medium was screened as optimal medium. The coincidence rate of positive results by ATP bioluminescence assay and by sterility test was 80%. However, one specimen identified as positive by ATP bioluminescence assay was judged as negative by sterility test, vice versa. Conclusion ATP bioluminescence assay was simple to handle and used widely, which was suitable for sterility test of intermediate product of vaccine.更多还原