【作者】 于霞； 李慧文； 姜广路； 张洪涛； 黄海荣；

【Author】 National Tuberculosis Reference Laboratory,Beijing Tuberculosis & Thoracic Tumor Institute, Beijing,China 101149 YuX,LiH,JiangG,ZhangH,HuangH~*

【机构】 国家结核病参比实验室北京市结核病胸部肿瘤研究所；

【摘要】 目的研究利福平储存液、含药罗氏培养基和7H9液体培养基中利福平的稳定性,探讨利福平储存液和利福平含药培养基的保质期。方法应用高压液相色谱仪连续监测在4℃和-20℃保存的利福平储存液至满1个月,连续监测7H9和罗氏培养基分别在4℃和37℃的储存条件下利福平浓度的变化至满6个月;通过两因素析因设计资料的方差分析来寻找利福平在罗氏培养基中不稳定的原因,比较不同的加热时间和加热温度对培养基中利福平浓度的影响;通过AlmarBlue指示剂筛选临床结核分枝杆菌分离株利福平的最低抑菌浓度(MIC)接近或低于1.0的菌株,监测菌株在应用不同存储时间的含利福平罗氏培养基上进行比例法药敏试验的结果变化。结果利福平储存液在4℃和-20℃保存的条件下,1个月内浓度未发生明显变化;在37℃保存的情况下,罗氏培养基和7H9液体培养基中的利福平分别在19天和6周后降低至可检测浓度范围下;在4℃保存的情况下,罗氏培养基和7H9培养基中利福平均降解约25%,6个月后罗氏培养基中的利福平降至可检测浓度以下,7H9中的利福平浓度仅为最初的37.5%;两因素析因设计资料的方差分析发现85℃加热50分钟导致20%的利福平降解,罗氏培养基中的蛋白成分和加热均可导致利福平降解,但加热的作用更明显;药敏结果显示,同一株MIC接近或低于耐药判断标准临界值的临床分离株在分别接种4℃存储0-6月的罗氏培养基中所获得的结果存在差异,当接种长时间保存的培养基时,发生敏感菌判断为耐药株的可能性大。结论利福平储存液至少在1个月内浓度保持稳定;含药罗氏培养基的保存时间应在2个月内;含药培养基的制作过程中应该严格控制加热的时间和加热温度。更多还原

【Abstract】 Objective To study the stability of Rifampicin(RFP) in Storage Solution,L-J media and 7H9 media, and investigate the shelf life of RFP storage solution and drug-containing media.Methods RFP in Storage Solution was monitored by HPLC for 1 month and the storage conditions were 4℃and -20℃.RFP decay in7H9 and L-J media was monitored serially by HPLC for 6 months and the storage conditions were 4℃and 37℃.Analysis of Variance for a two-way factorial design was used to find the causes of RFP instability in L-J media.Different heating time and heating temperature was compared to effect the concentration of RFP.Microplate Almar Blue array was used to determine the minimum inhibitory concentration(MIC) of clinical strains.Clinical strains(MIC<=1.0) was choose to receive the proportion method on different storage time(0-6month) of L-J media.Results RFP in Storage Solution whatever at 4℃or -20℃had no obvious change within 1 month.No RFP curve could be detected for L-J and 7H9 media after 3 weeks’ and 6 weeks’ incubation respectively at 37℃.25%decay was found both in 7H9 and L-J media at 4℃for 1 month.No RFP curve could be detected for L-J media and RFP in 7H9 media reduced to 37.5%after 6 months.Analysis of Variance for a two-way factorial design proved that concreting at 85℃for 50 min during the preparation of L-J media led about 25%RFP decay,heat and protein all led to RFP decay and the role of heat was more obvious than the protein.Drug susceptibility tests showed the same clinical isolate with MIC close to or below the resistance threshold had different results when inoculated on different storage time(0-6 months ) of L-J media at 4℃.When inoculated on prolonged storage time of L-J media,there was a possibility that sensitive strains would be judged to drug-resistant strains.Conclusions RFP storage solution was stable for at least 1 month.The storage time of drug-containing L-J medium should be no more than 2 months.In the preparation of the drug-containing media,the heating time and heating temperature should be strictly controlled.更多还原