期刊
JOURNAL OF VETERINARY SCIENCE
卷 15, 期 2, 页码 317-325出版社
KOREAN SOC VETERINARY SCIENCE
DOI: 10.4142/jvs.2014.15.2.317
关键词
cattle farm; E. coli O157; LAMP; shiga toxin; stx
资金
- National Research Foundation of Korea - Korean Government [2012R1A1A1012293]
- Research Settlement Fund for the new faculty of Seoul National University, Korea
- National Research Foundation of Korea [2012R1A1A1012293] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
A multiplex loop-mediated isothermal amplification (mLAMP) assay was developed for simultaneous detection of the stx1 and stx2 genes and applied for detection of shiga toxin-producing Escherichia colt (STEC) in cattle farm samples. Two target genes were distinguished based on T. values of 85.03 0.54 C for stx1 and 87.47 +/- 0.35 C for stx2. The mLAMP assay was specific (100% inclusivity and exclusivity), sensitive (with a detection limit as low as 10 fg/mu L), and quantifiable (R-2 = 0.9313). The efficacy and sensitivity were measured to evaluate applicability of the mLANIP assay to cattle farm samples. A total of 12 (12/253; 4.7%) and 17 (17/253; 6.7%) STEC O157, and 11 (11/236; 4.7%) non-O157 STEC strains were isolated from cattle farm samples by conventional selective culture, immunomagnetic separation, and PCR-based culture methods, respectively. The coinciding multiplex PCR and mLAMP results for the types of shiga toxin revealed the value of the mLAMP assay in terms of accuracy and rapidity for characterizing shiga toxin genes. Furthermore, the high detection rate of specific genes from enrichment broth samples indicates the potential utility of this assay as a screening method for detecting STEC in cattle farm samples.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据