Key words: Listeria monocytogenes, poultry, processing plant, evisceration. ABSTRACT . Listeria antisera (Denka Seiken, Tokyo, Japan), accord- ing to the . Caracterização feno e genotípica de cepas de Listeria monocytogenes isoladas They were also serotyped (Denka Seiken, Japan), and sub–typed by PFGE. Antisera for the typing of Listeria Group O and H antigens. Aids public health and food testing laboratories in identifying sources of infection/contamination to.
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The filters were then blocked as before, and the same procedure was performed, except that L. Similarly, to directly compare H-antigen determination by the ELISA method to that by the slide agglutination method, isolates initially were subcultured three times on BHI motility agar, as recommended in the slide agglutination kit protocol, to increase the proportion of fully flagellated cells and thereby render a robust reaction with the appropriate H-factor antisera.
It was observed that the spatial or temporally unrelated strains exhibited similar PFGE profiles, indicating a possible clonal relationship among them. Wells were washed twice with wash buffer 0.
The antimicrobial susceptibility of the isolates was evaluated by the microdilution broth method lisyeria ampicillin, gentamicin, trimethoprim, sulfamethoxazole and vancomycin. Genomic fingerprinting of 80 strains from the W. As an epidemiological tool, serotyping is not sufficiently discriminatory to determine positive correlations listedia food-borne isolates and clinical isolates in cases of listeriosis outbreaks, though a negative relationship between isolates could be demonstrated by differing serotype results in these instances Sometimes, such labs are not capacitated to make a correctly identification of this bacterium.
Acknowledgments We thank C. Current methods to subtype L.
Epidemiology of human listeriosis. We determined that the O-factor and H-factor antisera gave suitable results at dilutions of 1: Effectiveness of prevention efforts? However, this current favorable seilen of quasiuniform susceptibility to the antibiotics used in clinical practice may deteriorate under the selective pressure exerted by overuse of the drugs in the country. Because serotype reactions depend on the quality of the antisera used, which in turn depends on which standardized strains and antigen preparation methods are chosen, the likelihood of inaccurate or inconsistent assignment of serotype rises when laboratories prepare their own antisera, especially for nonclinical isolates i.
Nevertheless, these data were of particular concern since emergence and dissemination of antibiotic resistance in clinical isolates of L. All subsequent steps were performed at room temperature. The ELISA and colony immunoblotting techniques described in this paper are useful means to quickly and economically serotype L.
Correlations between molecular subtyping seuken serotyping of Listeria monocytogenes. The ELISA serotyping protocol described in this work is a means to attempt to rectify the ambiguities and intra- and interlaboratory variability inherent to the traditional serotyping method, while at the same time allowing laboratories lisferia perform more analyses with significant cost and time savings.
Listeriosis has been mainly reported by those countries, with few or even no reports from Africa, Asia and South America. Listeria monocytogenes in renal transplant recipients.
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In order for these risks to be minimized, subtyping of L. The standard slide agglutination method relies on visual acuity and judgement to produce accurate data; this may be a substantial source of variability when comparing data between individuals in a laboratory and between different laboratories.
Open in a separate window. The discrepancy was investigated by developing a serogroup-specific colony immunoblot method, which could distinguish these serogroups by differential staining Fig.
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Author information Article notes Copyright and License information Disclaimer. Isolates were confirmed as L. Survival of Listeria monocytogenes in soil. From January to Maythirteen isolates of L. We have adapted a commercially available set of L. Listeria monocytogenes is a gram-positive, food-borne bacterial pathogen that causes listeriosis in susceptible individuals 4. With the exception of a strain previously serotyped as 4e by agglutination, which was consistently serotyped as 4b by ELISA, strains representative of all serotypes were typeable by this method.
After color development, alkaline phosphatase was inactivated by incubating the filters for 10 min in 50 mM EDTA, pH 8. In Brazil, few clinical cases of listeriosis have been reported and none of them had their source of infection established. After digestion with ApaI and AscI, the strains were distributed in 3 different groups according to their profile.
These strains were isolated from various sources, including produce, meats and dairy products, food processing environments, animals and humans, soils, and environmental samples.
Unfortunately, in the present study the sources of exposure were undetermined for listerja the 12 clinical cases evaluated and no epidemiological data were also available, for this reason no conclusive relationships could be determined. How to cite this article. However, autoclaved cells losteria to react less to negative O-factor antisera relative to cells treated by the other methods; that is, lower background reactions seikne observed.
Incidence of Listeria monocytogenes in nature.