[全自动RAISUS使用早期收获的细胞悬浮液进行物种鉴定和抗菌敏感性试验]。

Isamu Nakasone, Kyoko Kisanuki, Miyako Higa, Tohru Kinjo, Nobuhisa Yamane
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引用次数: 0

摘要

我们评估了早期收获的细菌细胞悬浮液对全自动RAISUS (Nissui Pharmaceuticals Co., Ltd, Tokyo)的有用性,以便在原代培养物过夜孵育后一天内提供物种鉴定和抗菌药物敏感性测试结果。原代培养板上出现一个分离良好的菌落,将其转移到血琼脂或巧克力琼脂板上,然后孵育3至6小时。将早期收获的细菌细胞配制成McFarland 0.5浊度的RAISUS细胞悬液。对金黄色葡萄球菌ATCC 29213、粪肠球菌ATCC 29212、肺炎链球菌ATCC 49619、大肠杆菌ATCC 25922、铜绿假单胞菌ATCC 27853 5株ATCC参比菌株进行重复鉴定,鉴定结果均可接受。用上述5株菌株与流感嗜血杆菌ATCC 49247进行药敏试验。获得的结果表明,大多数药敏试验结果与标准试验程序获得的mic相当,但某些菌株,特别是流感嗜血杆菌和铜绿假单胞菌,对某些抗菌剂的mic存在显著差异。MIC测定结果的显著差异是由于制备的细胞悬浮液中活细胞浓度的差异。通过对实验室工作流程的分析,发现18S到20S的测试在下午5点前完成,而需要等到凌晨3点才能完成90S的测试。上述结果表明,早期收获的细菌细胞悬浮液可用于RAISUS菌种鉴定,但需调整活菌浓度以适应药敏试验。同时,为了提高RAISUS测试功能的快速性,迫切需要重构日常工作流程。
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[Species-identification and antimicrobial susceptibility tests by the fully automated RAISUS using an early-harvested cell suspension].

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.

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