Manufacture of an Automatic Test System Based on Standard Antifungal Agent Sensitivity Testing Metho

我们试制并评价了一套使用标准微肉汤稀释法进行抗真菌药敏试验的试剂盒。首先，为了确保该试剂盒能够方便地用于医疗机构的常规检测，我们确定了生产抗真菌剂结合的96孔微孔板的条件，可以冷冻储存。接下来，我们将冷冻板、无菌RPMI 1640培养基和无菌生理盐水制备真菌接种物的试剂盒分发给评估机构或合作评估机构。利用该试剂盒对新鲜临床分离的酵母样真菌进行了抗真菌药物敏感性试验。终点用目测法和平板分析仪测定。在质量控制试验中，标准真菌染色液采用平行试验。在日本的不同设施中，共测试了13种150株酵母样真菌菌株。实验结果表明，所制板的测定结果与常规标准法的测定结果具有良好的相关性。此外，设备的结果与研究人员重复测试的结果之间也获得了良好的相关性，证实了良好的可重复性。我们得出结论，该试剂盒在常规临床检测中产生了可靠的结果，表明它是一种非常有用的检测系统。基于这些结果，我们申请了该试剂盒作为诊断产品的生产许可，并获得了批准。

We have trial produced and evaluated a kit for performing antifungal agent sensitivity test using the satndard micro-broth dilution method. Firt, to ensure that the kit will be easily available for routine testing in medical facilities, we determined the condition for producing antifungal agent-incorporated 96-well microplate that can be stored frozen. Nex, we distributed the kits that contained the frozen plate, sterile RPMI 1640 medium for culture and sterile physiological saline for preparing fungal inoculum, to evaluating facilities or facilities cooperated in evaluation. Using these kits, antifungal agent sensitivity tests were conducted on fresh clinical isolates of yeast-like fungi. End-points were determined both visually and using a plate analyzer. Standard fungal stain was always tested in parallel during testing for quality control. A total of 150 yeast-like fungal strains belonging to 13 species were tested in various facilities in Japan. The showed a good correlation between the results obtained using the trial produced plates and those by conventional standard method. Furthmore, good correlation was also obtained between the results of the facilities and those of researchers repeating the tests, confirming good reproducibility. We concluded that this kit when used in routine clinical testing produced reliable results, showing that it is highly useful test system. Based on these results, we applied for permission to namufacture this kit as a diagnostic product, and obtained approval.