A real-time loop-mediated isothermal amplification method for rapid detection of Lawsonia intracellularis in porcine fecal samples

作者：Li, Yanan; Wang, Jianchang; Wang, Jinfeng; Liu, Libing; Zhang, Ruoxi; Shi, Ruihan; Han, Qingan; Sun, Jiguo; Yuan, Wanzhe^*

来源：Journal of Microbiological Methods, 2018, 151: 62-65.

DOI：10.1016/j.mimet.2018.06.002

• Lawsonia intracellularis
• 16S rRNA
• Real-time LAMP
• Visual inspection
• Molecular diagnosis

Porcine proliferative enteritis is a common diarrheal disease characterized by thickening of the intestinal mucosa in swine due to enterocyte proliferation, which is caused by Lawsonia intracellularis. In this study, a real-time loop-mediated isothermal amplification (LAMP) assay was developed to detect L. intracellularis based on the conserved region of the 16S ribosomal RNA gene. The optimal reaction conditions of the real-time LAMP was 65 degrees C for 60 min. The LAMP products could be detected by both real-time turbidity and direct visual inspection. The assay was specific for L. intracellularis, as no cross-reaction was observed with other pathogens. The detection limit of the real-time LAMP assay was 1.4 x 10(-)(1)pg of L. intracellularis DNA, which was the same as that of real-time PCR and approximately 100 times more sensitive than that of conventional PCR. Of the 136 clinical samples, L. intracellularis DNA was identified in 60 samples by real-time LAMP, which was the same as real-time PCR and higher than conventional PCR (36.8%, 50/136). The specific, sensitive and rapid real-time LAMP assay developed in this study could be a useful alternative tool in point-of-care (POC) diagnosis of L. intracellularis infection.

• 出版日期2018-8
• 单位河北农业大学