RAPID DETECTION OF BRUCELLA ABORTUS BY A NOVEL PROXIMITY LIGATION‐BASED LOOP‐MEDIATED ISOTHERMAL AMPLIFICATION METHOD

Abstract

ABSTRACT Brucellosis is one of the most common zoonotic diseases, and current methods of detecting this pathogen are quite difficult. This work combines the benefits of a proximity ligation assay with those of a loop‐mediated isothermal amplification method to develop a novel proximity ligation‐based loop‐mediated isothermal amplification method useful for Brucella detection. The genomic DNA extraction procedure is not needed. Sensitivity of this assay for detecting Brucella abortus is 1 × 104 cells/mL in buffer and 1 × 105 cells/mL in milk. The time to detection is within 2 h of initiating the procedure, and no special equipment is needed. This new method is also suitable for the detection of other pathogens, and as such will be useful in the food safety industry.PRACTICAL APPLICATIONSPolymerase chain reaction (PCR) is a sensitivity method for microbe detection, but the complicated genomic DNA extraction procedure and costly equipment needed for this method makes the PCR method unpopular in developing countries. In this study, we present the novel proximity ligation‐based loop‐mediated isothermal amplification (P‐LAMP) method for Brucella detection; this is the first time to combine the monoclonal antibody for identify microbe and LAMP method for high performance amplification DNA. The genomic DNA extraction procedure is not needed and a water‐bath boiler is the only equipment required to complete the detection process. The P‐LAMP method is useful for food safety pathogen detection in developing countries.