Loop-mediated isothermal amplification assay forEnterococcus sp., E. coliandS. aureusin chicken

Abstract

Bacterial chondronecrosis with osteomyelitis (BCO) is a major cause of lameness in broiler chicken, and results in serious economic losses worldwide. Although the pathogenesis mechanism leading to lameness is not entirely understood, some strains of Enterococcus sp., avian pathogenic Escherichia coli, or Staphylococcus aureus have been long recognized as important causative pathogens. To prevent the progression of Enterococcus sp., avian pathogenic E. coli, or S. aureus infections, we developed rapid, sensitive, and convenient diagnostic assays using loop-mediated isothermal amplification (LAMP). Entero-Common-LAMP assays were developed for a simultaneous detection of eight Enterococcus species. To target specific microorganisms, seven Entero-Specific-LAMP assays for E. faecalis, E. faecium, E. hirae, E. gallinarum, E. avium, E. durans and E. cecorum, and E. coli-LAMP and S. aureus-LAMP assays, were developed. Considering the prevalence and economic impact of Enterococcus sp., E. coli, and S. aureus, the developed ten different LAMP assays have a considerable potential as routine diagnostic methods in the field or in resource-limited environments.

A Microfluidic Chip for LAMP-based Multiplex Detection of Pathogen

“…The total genomic DNA was extracted by using kit, following the instruction from manufacturer. The targets gene used for these bacterial detections were ecolc3109_1 [18], nuc [19]and opr [20],…”