Quantitative determination of gelatinase activity among enterococci.
Gelatinase, hemolysin and aggregation substance have all been reported to be virulence factors of enterococci. In this study, gelatinase production was investigated in isolates of Enterococcus faecalis (n=93), E. faecium (n=49) and E. avium (n=36) recovered from hospitalized patients. Gelatinase was detected in 45% of E. faecalis isolates, but could not be detected in E. faecium and E. avium. Gelatinase activity was then measured by radial diffusion for the 42 gelatinase-positive E. faecalis isolates. To convert gelatinase activity into proteinase K activity, a standard curve was produced by placing different concentrations of proteinase K into wells in the gelatine plate. Gelatinase activity per E. faecalis colony ranged from 2.6 x 10(-7) to 2.2 x 10(-5) microg/ml, proportionate to the activity of proteinase K. An approximately 84-fold difference in gelatinase concentration was observed between the colony producing the highest amount and that producing the lowest amount. This method may be useful for determining the virulence of given isolates in relation to gelatinase production as it is quick, easy and inexpensive to perform.
Kanemitsu
K
K
Department of Molecular Diagnostics, Tohoku University Graduate School of Medicine, 1-1 Seiryocho Aobaku Sendai, Miyagi 980-8574, Japan. kane2@mail.cc.tohoku.ac.jp
Nishino
T
T
Kunishima
H
H
Okamura
N
N
Takemura
H
H
Yamamoto
H
H
Kaku
M
M
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001431:Bacteriological Techniques; D003470:Culture Media; D004058:Diffusion; D016983:Enterococcus / Q000201:enzymology* / Q000472:pathogenicity; D013293:Enterococcus faecalis / Q000201:enzymology / Q000472:pathogenicity; D016984:Enterococcus faecium / Q000201:enzymology / Q000472:pathogenicity; D005260:Female; D005780:Gelatin / Q000378:metabolism; D018093:Gelatinases / Q000378:metabolism*; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology*; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged
D003470:Culture Media; D005780:Gelatin; D018093:Gelatinases