RRC ID 2709
著者 Selim AS, Boonkumklao P, Sone T, Assavanig A, Wada M, Yokota A.
タイトル Development and assessment of a real-time pcr assay for rapid and sensitive detection of a novel thermotolerant bacterium, Lactobacillus thermotolerans, in chicken feces.
ジャーナル Appl Environ Microbiol
Abstract A new real-time PCR assay was successfully developed using a TaqMan fluorescence probe for specific detection and enumeration of a novel bacterium, Lactobacillus thermotolerans, in chicken feces. The specific primers and probe were designed based on the L. thermotolerans 16S rRNA gene sequences, and these sequences were compared to those of all available 16S rRNA genes in the GenBank database. The assay, targeting 16S rRNA gene, was evaluated using DNA from a pure culture of L. thermotolerans, DNA from the closely related bacteria Lactobacillus mucosae DSM 13345(T) and Lactobacillus fermentum JCM 1173(T), and DNA from other lactic acid bacteria in quantitative experiments. Serial dilutions of L. thermotolerans DNA were used as external standards for calibration. The minimum detection limit of this technique was 1.84 x 10(3) cells/ml of an L. thermotolerans pure culture. The assay was then applied to chicken feces in two different trials. In the first trial, the cell population was 10(4) cells/g feces on day 4 and 10(5) cells/g feces on days 11 to 18. However, cell populations of 10(6) to 10(7) cells/g feces were detected in the second trial. The total bacterial count, measured by 4',6-diamidino-2-phenylindole (DAPI) staining, was approximately 10(11) cells/g feces. These results suggest that in general, L. thermotolerans is a normal member of the chicken gut microbiota, although it is present at relatively low levels in the feces.
巻・号 71(8)
ページ 4214-9
公開日 2005-8-1
DOI 10.1128/AEM.71.8.4214-4219.2005
PII 71/8/4214
PMID 16085805
PMC PMC1183300
MeSH Animals Chickens / microbiology* Colony Count, Microbial DNA Primers DNA, Bacterial / analysis Feces / microbiology* Hot Temperature* Lactobacillus / classification Lactobacillus / genetics Lactobacillus / growth & development* Lactobacillus / isolation & purification* Polymerase Chain Reaction / methods* Species Specificity Time Factors
IF 4.016
引用数 18
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY
リソース情報
病原微生物 JCM 11425? JCM 1044? JCM 5667? JCM 5666? JCM 5810? JCM 2011? JCM 1126? JCM 1134? JCM 1120? JCM 1173? JCM 1132? JCM 7876? JCM 1158? JCM 8724? JCM 5887T? JCM 1564? JCM 1395? JCM 1391? JCM 3817? JCM 2257?