利用环介导的等温扩增法快速灵敏地检测针对hilA基因的沙门氏菌。

Q2 Agricultural and Biological Sciences Genomics and Informatics Pub Date : 2021-09-01 Epub Date: 2021-09-30 DOI:10.5808/gi.21048
Jiyon Chu, Juyoun Shin, Shinseok Kang, Sun Shin, Yeun-Jun Chung
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引用次数: 5

摘要

沙门氏菌是引起食源性疾病暴发的主要病原体之一。在这项研究中,我们旨在建立一种环介导的等温扩增(LAMP)方法来快速灵敏地检测沙门氏菌。我们设计了针对hilA基因的LAMP引物,作为沙门氏菌的通用标记。本研究共使用了7株沙门菌和11株非沙门菌病原菌,分别来自8个属。所有沙门氏菌LAMP扩增结果均为阳性;而对非沙门菌无非特异性扩增信号。检测限为100飞图(每个反应20拷贝),灵敏度是传统PCR检测限(100 pg)的1000倍。阳性扩增信号的反应时间不到20分钟,不到传统PCR所用时间的三分之一。综上所述,与传统的PCR方法相比,LAMP法能够准确地检测出沙门氏菌种类,灵敏度更高,速度更快,可用于现场即时检测。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Rapid and sensitive detection of Salmonella species targeting the hilA gene using a loop-mediated isothermal amplification assay.

Salmonella species are among the major pathogens that cause foodborne illness outbreaks. In this study, we aimed to develop a loop-mediated isothermal amplification (LAMP) assay for the rapid and sensitive detection of Salmonella species. We designed LAMP primers targeting the hilA gene as a universal marker of Salmonella species. A total of seven Salmonella species strains and 11 non-Salmonella pathogen strains from eight different genera were used in this study. All Salmonella strains showed positive amplification signals with the Salmonella LAMP assay; however, there was no non-specific amplification signal for the non-Salmonella strains. The detection limit was 100 femtograms (20 copies per reaction), which was ~1,000 times more sensitive than the detection limits of the conventional polymerase chain reaction (PCR) assay (100 pg). The reaction time for a positive amplification signal was less than 20 minutes, which was less than one-third the time taken while using conventional PCR. In conclusion, our Salmonella LAMP assay accurately detected Salmonella species with a higher degree of sensitivity and greater rapidity than the conventional PCR assay, and it may be suitable for point-of-care testing in the field.

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来源期刊
Genomics and Informatics
Genomics and Informatics Agricultural and Biological Sciences-Ecology, Evolution, Behavior and Systematics
CiteScore
1.90
自引率
0.00%
发文量
0
审稿时长
12 weeks
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