Loop-mediated isothermal amplification for the rapid, sensitive, and specific detection of the O9 group of Salmonella

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• 作者：Masashi Okamura ; Yousuke Ohba ; Shuichi Kikuchi ; Akiko Suzuki ; Hajime Tachizaki ; Kazuaki Takehara ; Masanari Ikedo ; Tadashi Kojima ; Masayuki Nakamura
• 关键词：Loop-mediated isothermal amplification (LAMP) ; Salmonella enterica serovar Enteritidis ; Chicken
• 刊名：Veterinary Microbiology
• 出版年：2008
• 出版时间：25 November 2008
• 年：2008
• 卷：132
• 期：1-2
• 页码：197-204
• 全文大小：252 K

文摘

In the present study, the loop-mediated isothermal amplification (LAMP) assay was developed to amplify the fragments of the O9 Salmonella-specific insertion element and evaluated in the laboratory for its potential use in a field situation, such as poultry farms. Among the bacteria tested, a positive reaction was observed only for 128 strains of 6 serovars of the O9 group Salmonella, such as Enteritidis (SE) and Pullorum. The detection limit of the LAMP assay was 10³ CFU/ml, which was more sensitive than that of the polymerase chain reaction (PCR) assay with the same target gene (10⁶ CFU/ml). The final results were obtained within 30 min for the LAMP assay, while the PCR assay needed a total of 120 min. When the LAMP assay was applied to the enrichment broth mixed with cecal dropping samples either spiked with SE in vitro or excreted by SE-inoculated hens, the results were comparable to those of the conventional plating method including 2 separate enrichments. In conclusion, the LAMP assay developed in the present study is an effective method for the specific detection of the O9 group Salmonella serovars, including SE.