title: Molecular Serogrouping of Listeria monocytogenes from Brazil Using PCR.
abstract: We assessed the serotype distribution of Listeria monocytogenes isolates from clinical, beef, and environment samples using two PCR-based protocols for serogrouping. A panel of 134 isolates (22 clinical samples, 79 samples of beef cuts, and 33 samples from the beef processing environment) were subjected to conventional serology and identified as serotypes 1/2a (n = 12), 1/2b (n = 21), 1/2c (n = 71), and 4b (n = 30). Isolates from clinical samples were predominantly serotype 4b, and the most prevalent serotype among the beef cut and environment samples was 1/2c. The protocol described by M. Doumith, C. Buchrieser, P. Glaser, C. Jacquet, and P. Martin (J. Clin. Microbiol. 42:3819-3822, 2004) produced contradictory results for seven 1/2a isolates, which were positive for lmo1118 and had the profile IIc (serotypes 1/2c and 3c). Fifteen serotype 4b isolates amplified the target lmo0737, with the atypical profile IVb variant 1. The results obtained with the protocol described by M. K. Borucki and D. R. Call (J. Clin. Microbiol. 41:5537-5540, 2003) were in full agreement with those of the conventional serology. We recommend using this multiplex PCR approach by adding one pair of the reported primers to the panel to reduce total effort by one PCR while maintaining specificity. We present additional recommendations to improve the efficiency and reproducibility of this serogrouping assay.
authors.affiliation: Universidade Federal de Viçosa, Departamento de Veterinária, Viçosa, MG, Brazil.
authors.forename: Anderson Carlos
authors.initials: AC
authors.lastname: Camargo
authors.affiliation: Fundação Oswaldo Cruz, Instituto Oswaldo Cruz, Laboratório de Zoonoses Bacterianas, Rio de Janeiro, RJ, Brazil.
authors.forename: Deyse Christina
authors.initials: DC
authors.lastname: Vallim
authors.affiliation: Fundação Oswaldo Cruz, Instituto Oswaldo Cruz, Laboratório de Zoonoses Bacterianas, Rio de Janeiro, RJ, Brazil.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Hofer
authors.affiliation: Universidade Federal de Viçosa, Departamento de Veterinária, Viçosa, MG, Brazil. nero@ufv.br.
authors.forename: Luís Augusto
authors.initials: LA
authors.lastname: Nero
mesh_terms: D001938:Brazil; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D006801:Humans; D008089:Listeria monocytogenes / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008088:Listeriosis / Q000382:microbiology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012703:Serotyping / Q000379:methods*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial