Establishment of Three Rapid Visual Detection Methods for Burkholderia gladioli pv. cocovenenans Based on Body Temperature Amplification

Abstract

Three rapid visual methods, namely chromogenic, fluorescence and test strip, for the rapid detection of Burkholderia gladioli pv. cocovenenans in foods were established based on enzymatic recombinase amplification (ERA). Primers and probes were designed and screened based on the bonM gene of B. gladioli pv. cocovenenans. Then the specificity and sensitivity of the three methods were evaluated, as well as their applicability and accuracy in the detection of commercial food samples. The results showed that three strains of B. gladioli pv. cocovenenans, but not other common foodborne pathogens and other B. gladioli strains, were amplified by the three methods, indicating their good specificity. The detection limits of these methods were all 10-2 ng/μL, and their sensitivity was good. Out of 15 commercial samples, two tested positive by each of these methods with a detection rate of 13.3%. This result was consistent with that of the national standard method, indicating that our methods had good applicability and accuracy. All three methods give results that can be observed by the naked eye after amplification at 37 ℃ for 15 min, which provide a new and simple strategy for the rapid, visual and on-site screening of B. gladioli pv. cocovenenans in foods.

Keywords

• burkholderia gladioli pv. cocovenenans; enzymatic recombinase amplification; visualization; rapid detection