Rapid method for the differentiation of gram-positive and gram-negative bacteria on membrane filters.

Abstract:

:Microfiltration has become a popular procedure for the concentration and enumeration of bacteria. We developed a rapid and sensitive method for the differentiation of gram-positive and gram-negative bacteria, utilizing a polycarbonate membrane filter, crystal violet, iodine, 95% ethanol, and 6% carbol fuchsin, that can be completed in 60 to 90 s. Gram reactions of 49 species belonging to 30 genera of bacteria were correctly determined by the filter-Gram stain. The sensitivities of the filter-Gram stain and conventional slide-Gram stain were compared by testing dilutions of Escherichia coli, Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae suspensions in the presence and absence of whole human blood. The filter-Gram stain was approximately 100-fold more sensitive than the slide-Gram stain. The filter-Gram stain detected 2 to 100 bacteria, whereas the slide-Gram stain failed to detect less than 1,000 bacteria. The sensitivities of the methods were not significantly altered by the addition of whole human blood to the dilutions of bacteria tested. The filter-Gram stain could be a useful tool for the examination of body fluids with very low numbers of bacteria.

journal_name

J Clin Microbiol

authors

Romero S,Schell RF,Pennell DR

doi

10.1128/JCM.26.7.1378-1382.1988

subject

Has Abstract

pub_date

1988-07-01 00:00:00

pages

1378-82

issue

7

eissn

0095-1137

issn

1098-660X

journal_volume

26

pub_type

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