柠檬醛体外诱导白念珠菌耐药实验探究.docVIP

柠檬醛体外诱导白念珠菌耐药实验探究 　 作者：张文平，王小丽， 黄真，曹镐禄 【关键词】 白色念珠菌；,,柠檬醛；,,氟康唑；,,耐药 　　摘要：目的探讨柠檬醛体外诱导白念珠菌耐药性的反应情况，进一步评价柠檬醛的临床应用价值。方法①参照NCCLS M27A方案中的微量液基稀释法测定柠檬醛和氟康唑对原始白念珠菌的最低抑菌浓度(MIC)和最低杀菌浓度(MFC)；②采用多步诱导法，观察在含梯度浓度(1～50MIC)柠檬醛培养基上连续转种的白念珠菌MIC的变化情况，以氟康唑作为阳性对照，同时设空白对照和溶媒对照；③将敏感性下降的诱导株在无药培养基上连续转种10次后，检测其MIC值；④测定柠檬醛对氟康唑敏感性下降诱导株的MIC值。结果①柠檬醛和氟康唑对原始白念珠菌的MIC值分别为：0.162 μg/ml和2.0 μg/ml；柠檬醛对原始白念珠菌的MFC值为0.324 μg/ml，氟康唑在实验的最高浓度(64 μg/ml)未发现有杀菌作用。 ②在多步诱导培养过程中，柠檬醛组的MIC值始终保持不变，未诱导出对柠檬醛耐药的白念珠菌菌株，菌株转种至含10 MIC柠檬醛的培养基时，即停止生长，而在含50 MIC氟康唑的培养基上菌株仍能生长。氟康唑诱导组菌株在培养过程中，MIC值逐渐升高，转种至50 MIC时，其MIC (命名为FR株)是其亲株的12倍。③FR在无药培养基上连续转种10次后得到FR＇，氟康唑在FR＇的MIC为20.16 μg/m，与FR相比，差异无显著性(P＞0.05)；④柠檬醛对FR的MIC值为0.189 μg/ml，与其相应的原始菌株无差异。结论 柠檬醛不易产生耐药性，且对氟康唑敏感性下降的诱导株仍敏感。柠檬醛是一种有良好开发前景的抗系统性念珠菌感染的天然药物。 　　关键词：白色念珠菌； 柠檬醛； 氟康唑； 耐药 　　Studies on Resistant Induction of Citral to Candida albicans in vitro 　　Abstract：ObjectiveTo investigate whether it is likely to induce Citral-resistant Candida albicans mutations in vitro, in order to give further evaluations of its antifungal actions for clinical use. Methods①The minimum inhibitory concentration(MIC) and the minimum fungicidal concentration(MFC) of citral against C. albicans were measured by broth microdilution method according to document M27-A published by the National Committee for Clinical Laboratory Standards(NCCLS); ② A C. albicans strain was induced by serial subcultures on agar medium containing different concentrations of Citral (equivalent to 1MIC～50MIC), and 3 groups were set as control groups which were fluconazole group (positive control), 1.5% alcohol group(solvent control), drug-free group, respectively. The susceptibilities (MIC) of this parental strain and its respective derivatives to Citral and fluconazole were measured. ③ The induced less susceptible derivatives were subcultured on drug-free medium for 10 times consecutively, then their MICs were tested. ④ The MIC of induced fluconazole less susceptible derivative to Citral was also measured. Results① MIC of citral and f