Nakasone Isamu, Kisanuki Kyoko, Higa Miyako, Kinjo Tohru, Yamane Nobuhisa
Clinical Laboratories, University Hospital of the Ryukyus.
Rinsho Biseibutshu Jinsoku Shindan Kenkyukai Shi. 2006;17(2):125-32.
We evaluated the usefulness of an early-harvested bacterial cell suspension to the fully automated RAISUS (Nissui Pharmaceuticals Co., Ltd., Tokyo) to provide the results of species-identification and antimicrobial susceptibility testings within a day after overnight-incubation of the primary cultures. A single, well-separated colony appeared on the primary culture plate was transferred onto a blood agar or chocolate agar plates, then incubated for 3 to 6 hours. The cell suspension to the RAISUS was properly prepared to the McFarland 0.5 turbidity from the early-harvested bacterial cells. When the five ATCC reference strains, consisting of Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212, Streptococcus pneumoniae ATCC 49619, Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853, were repeatedly tested for the species-identification, all the identification results were acceptable. Antimicrobial susceptibility tests were evaluated with the above five strains and Haemophilus influenzae ATCC 49247. The results obtained indicated that the most susceptibility test results were comparable to those MICs obtained by the standard test procedure, but some strains, in particular, H. influenzae and P. aeruginosa gave significantly discrepant MICs for certain antimicrobial agents. The significant discrepancy in MIC determinations regarded the difference of viable cell concentrations in the cell suspension prepared respectively. Through the analysis of laboratory workflow, it became to apparent that 18S to 20S of the tests were completed by 5:00 p.m., and it required to wait until 3:00 a.m. to complete 90S of the tests. With these results, the early-harvested bacterial cell suspension is applicable to species-identification by RAISUS, but it is necessary to adjust viable cell concentrations to antimicrobial susceptibility test. Also, it is urgent to reconstitute a daily workflow to improve the rapidity of RAISUS test function.
我们评估了早期收获的细菌细胞悬液对于全自动RAISUS(日本东京日水制药株式会社)的实用性,该仪器能够在初代培养过夜培养后一天内提供菌种鉴定和抗菌药敏试验结果。将初代培养平板上单个、分离良好的菌落转移至血琼脂或巧克力琼脂平板上,然后培养3至6小时。从早期收获的细菌细胞制备出符合麦氏0.5浊度的RAISUS细胞悬液。当对由金黄色葡萄球菌ATCC 29213、粪肠球菌ATCC 29212、肺炎链球菌ATCC 49619、大肠埃希菌ATCC 25922和铜绿假单胞菌ATCC 27853组成的5株ATCC参考菌株进行多次菌种鉴定时,所有鉴定结果均可接受。使用上述5株菌株和流感嗜血杆菌ATCC 49247进行抗菌药敏试验评估。所得结果表明,大多数药敏试验结果与通过标准试验程序获得的最低抑菌浓度(MIC)结果相当,但某些菌株,特别是流感嗜血杆菌和铜绿假单胞菌,对于某些抗菌药物的MIC值存在显著差异。MIC测定中的显著差异是由于分别制备的细胞悬液中活细胞浓度不同所致。通过对实验室工作流程的分析,明显发现18S至20S的试验在下午5:00完成,而完成90S的试验则需要等到凌晨3:00。基于这些结果,早期收获的细菌细胞悬液适用于RAISUS进行菌种鉴定,但进行抗菌药敏试验时需要调整活细胞浓度。此外,迫切需要重新构建日常工作流程以提高RAISUS试验功能的速度。