Establishment of loop-mediated isothermal amplification for Brucella detection using a warmer pad as a heating source

Author:

Yin-Bo Ye1,Dong-Liang Li2,Bo-Han Yang1,Zhao Zhang1,Si-Yao Mei1,Bao-Shan Liu1ORCID,Ze-Liang Chen13

Affiliation:

1. Key Laboratory of Livestock Infectious Diseases, Ministry of Education, Shenyang Agricultural University, Shenyang City, Liaoning Province, 110866, China

2. Beijing Animal Disease Prevention & Control Center, Beijing, 102609, China

3. KeyLaboratory of Zoonose Prevention and Control at Universities of Inner Mongolia Autonomous Region, Innovative Institute of Zoonoses, Medical College, Inner Mongolia Minzu University, Tongliao, China

Abstract

The study sought to establish a sensitive and specific on-site loop-mediated isothermal amplification (LAMP) for Brucella heated using a warmer pad. LAMP primers specific to the conserved BvrR gene were designed, and the LAMP reaction was optimized. The heating characteristics of the warmer pad were investigated. The detection validity (specificity, sensitivity) of clinical samples by warmer-pad LAMP (WP-LAMP) was compared with that of qPCR. The WP-LAMP method displayed high specificity and sensitivity for five Brucella gene copies. The detection of 104 clinical samples was 97.1% concordant with quantitative polymerase chain reaction. The results showed the success of the WP-LAMP for on-site detection. The method requires no special equipment and is conducive to the prevention and control of brucellosis.

Funder

the State Key Program of National Natural Science of China

NSFC International (regional) cooperation and exchange program

Publisher

Future Science Ltd

Subject

General Biochemistry, Genetics and Molecular Biology,Biotechnology

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