rapid discrimination of gram-positive and gram-negative bacteria in liquid samples by using naoh-sodium dodecyl sulfate solution and flow cytometry快速歧视对革兰氏阳性和革兰氏阴性菌在液体样品通过naoh-sodium十二烷基硫酸溶液和流式细胞术.pdfVIP

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2017-09-09 发布于上海
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rapid discrimination of gram-positive and gram-negative bacteria in liquid samples by using naoh-sodium dodecyl sulfate solution and flow cytometry快速歧视对革兰氏阳性和革兰氏阴性菌在液体样品通过naoh-sodium十二烷基硫酸溶液和流式细胞术.pdf

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rapid discrimination of gram-positive and gram-negative bacteria in liquid samples by using naoh-sodium dodecyl sulfate solution and flow cytometry快速歧视对革兰氏阳性和革兰氏阴性菌在液体样品通过naoh-sodium十二烷基硫酸溶液和流式细胞术

Rapid Discrimination of Gram-Positive and Gram- Negative Bacteria in Liquid Samples by Using NaOH- Sodium Dodecyl Sulfate Solution and Flow Cytometry Atsushi Wada*, Mari Kono, Sawako Kawauchi, Yuri Takagi, Takashi Morikawa, Kunihiro Funakoshi Cell Analysis Center, Scientific Affairs, Sysmex Corporation, Nishi-ku, Kobe, Hyogo, Japan Abstract Background: For precise diagnosis of urinary tract infections (UTI), and selection of the appropriate prescriptions for their treatment, we explored a simple and rapid method of discriminating gram-positive and gram-negative bacteria in liquid samples. Methodology/Principal Findings: We employed the NaOH-sodium dodecyl sulfate (SDS) solution conventionally used for plasmid extraction from Escherichia coli and the automated urine particle analyzer UF-1000i (Sysmex Corporation) for our novel method. The NaOH-SDS solution was used to determine differences in the cell wall structures between gram-positive and gram-negative bacteria, since the tolerance to such chemicals reflects the thickness and structural differences of bacterial cell walls. The UF-1000i instrument was used as a quantitative bacterial counter. We found that gram-negative bacteria, including E. coli, in liquid culture could easily be lysed by direct addition of equal volumes of NaOH-SDS solution. In contrast, Enterococcus faecalis, which is a gram-positive bacterium, could not be completely lysed by the solution. We then optimized the reaction time of the NaOH-SDS treatment at room temperature by using 3 gram-positive and 4 gram- negative bacterial strains and determined that the optimum reaction time was 5 min. Finally, in order to evaluate the generalizability of this method, we treated 8 gram-positive strains and 8 gram-negative strains, or 4 gram-positive and 4 gram-negative strains incubated in voluntary urine from hea