关键词: 唐菖蒲伯克霍尔德氏菌椰毒致病变种, 酶促等温扩增技术, 可视化, 快速检测

Abstract: [Objective] In order to realize the rapid detection of Burkholderia gladioli pathovar cocoenans in food, based on Enzymatic Recombinase Amplification (ERA), this study established three visual and rapid detection methods for Burkholderia gladioli pathovar cocoenan. [Methods] The primers and probes of ERA were designed and screened according to the bonM gene of Burkholderia gladioli pathovar cocoenans to establish the three visual rapid detection methods: chromogenic method, fluorescence method and test strip method. Then the specificity and sensitivity of three methods were evaluated, as well as its applicability and accuracy in commercial food samples. [Results] The results showed that 3 strains of Burkholderia gladioli pathovar cocoenans were positive, common foodborne pathogens and other Burkholderia gladioli strains were negative, which showed that the established methods had good specificity. The detection limits of the three methods were all 10-2 ng/μL, and the sensitivity was good. 15 commercial samples were tested by three visual rapid detection methods. The results showed that two samples were positive, and the detection rate was 13.3%. The results were consistent with those of the national standard method, indicating that the methods had good applicability and accuracy. [Conclusion] The three visual rapid detection methods established in this study based on body temperature amplification technology, namely chromogenic method, fluorescence method and test strip method, have high specificity and sensitivity. 15 min of amplification at 37℃ can obtain the results and be seen with naked eyes. This provides a new and simple strategy of visual rapid detection on-site for Burkholderia gladioli pathovar cocovenenans in food.

Key words: Burkholderia gladioli pathovar cocovenenans, enzymatic Recombinase Amplification, visualization, rapid detection