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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

Wada, Atsushi ; Kono, Mari ; Kawauchi, Sawako ; Takagi, Yuri ; Morikawa, Takashi ; Funakoshi, Kunihiro Heimesaat, Markus M.

PloS one, 2012-10, Vol.7 (10), p.e47093 [Periódico revisado por pares]

United States: Public Library of Science

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Título:
Rapid discrimination of Gram-positive and Gram-negative bacteria in liquid samples by using NaOH-sodium dodecyl sulfate solution and flow cytometry
Autor: Wada, Atsushi ; Kono, Mari ; Kawauchi, Sawako ; Takagi, Yuri ; Morikawa, Takashi ; Funakoshi, Kunihiro
Heimesaat, Markus M.
Assuntos: Antibiotics ; Automation ; Bacteria ; Bacterial Infections - diagnosis ; Bacteriuria ; Biology ; Cell culture ; Cell walls ; Culture media ; Cytometry ; Deoxyribonucleic acid ; Diagnosis ; DNA ; E coli ; Enterococcus faecalis ; Enterococcus faecalis - cytology ; Enterococcus faecalis - isolation & purification ; Enterococcus faecalis - metabolism ; Escherichia coli ; Escherichia coli - cytology ; Escherichia coli - isolation & purification ; Escherichia coli - metabolism ; Flow cytometry ; Flow Cytometry - economics ; Flow Cytometry - methods ; Gram-negative bacteria ; Gram-Negative Bacteria - cytology ; Gram-Negative Bacteria - isolation & purification ; Gram-Negative Bacteria - metabolism ; Gram-positive bacteria ; Gram-Positive Bacteria - cytology ; Gram-Positive Bacteria - isolation & purification ; Gram-Positive Bacteria - metabolism ; Humans ; Infectious diseases ; Laboratories ; Liquid culture ; Medical diagnosis ; Medicine ; Methods ; Microbiology ; Microscopy ; Reaction time ; Sodium ; Sodium dodecyl sulfate ; Sodium Dodecyl Sulfate - metabolism ; Sodium Hydroxide - metabolism ; Sodium lauryl sulfate ; Staphylococcus ; Strains (organisms) ; Sulfates ; Thickness ; Time Factors ; Urinary tract ; Urinary tract diseases ; Urinary tract infections ; Urine ; Urine - microbiology ; Urine culture ; Urogenital system
É parte de: PloS one, 2012-10, Vol.7 (10), p.e47093
Notas: Conceived and designed the experiments: AW TM KF. Performed the experiments: AW. Analyzed the data: AW. Contributed reagents/materials/analysis tools: MK SK YT. Wrote the paper: AW.
Competing Interests: All the authors are employees of Sysmex Corporation, which is the company that produces the automated urine particle analyzer UF-1000i. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials.
Descrição: 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. 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 healthy volunteers in the same way and demonstrated that all the gram-positive bacteria were discriminated quantitatively from gram negative bacteria using this method. Using our new method, we could easily discriminate gram-positive and gram-negative bacteria in liquid culture media within 10 min. This simple and rapid method may be useful for determining the treatment course of patients with UTIs, especially for those without a prior history of UTIs. The method may be easily applied in order to obtain additional information for clinical prescriptions from bacteriuria.
Editor: United States: Public Library of Science
Idioma: Inglês

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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

Wada, Atsushi ; Kono, Mari ; Kawauchi, Sawako ; Takagi, Yuri ; Morikawa, Takashi ; Funakoshi, Kunihiro Heimesaat, Markus M.

United States: Public Library of Science

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