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Development and assessment of a Real-Time PCR assay for the rapid and sensitive detection of a novel thermotolerant bacterium, Lactobacillus thermotolerans, in chicken feces

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Title: Development and assessment of a Real-Time PCR assay for the rapid and sensitive detection of a novel thermotolerant bacterium, Lactobacillus thermotolerans, in chicken feces
Authors: Selim, Abu Sadeque Md. Browse this author
Assavanig, Apinya Browse this author
Sone, Teruo Browse this author →KAKEN DB
Boonkumklao, Piyanuch Browse this author
Wada, Masaru Browse this author
Yokota, Atsushi6 Browse this author →KAKEN DB
Authors(alt): 横田, 篤6
Issue Date: Aug-2005
Publisher: American Society for Microbiology
Journal Title: Applied and Environmental Microbiology
Volume: 71
Issue: 8
Start Page: 4214
End Page: 4219
Publisher DOI: 10.1128/AEM.71.8.4214-4219.2005
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 13345T and Lactobacillus fermentum JCM 1173T, 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 103 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 104 cells/g feces on day 4 and 105 cells/g feces on days 11 to 18. However, cell populations of 106 to 107 cells/g feces were detected in the second trial. The total bacterial count, measured by 4',6-diamidino-2-phenylindole (DAPI) staining, was approximately 1011 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.
Rights: Copyright © 2005 American Society for Microbiology
Type: article (author version)
URI: http://hdl.handle.net/2115/4863
Appears in Collections:農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 横田 篤

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