Sensitivity comparison of the ISE method and classical PCR to detect Burkholderia glumae

LUO Jin-yan;CHEN Lei;QIN Meng;ZHAO Yu-qiang;YU Hui;Shanghai City Agricultural Technology Extension and Service Center;National AgroTechnical Extension and Service Centre;

ISE;;PCR;;rice;;Burkholderia glumae

The present study combined the fluoride ion-selective electrode( ISE) method with classical PCR to increase the detection efficiency. The 4 antisera of rabbit anti whole cell of B. glumae have been successfully obtained by common serological method,which were ASBg14,ASBg32,ASBg26 and ASBg04. The antibody of 4 antisera could be effective when diluted by 5 120 times and reached 1∶ 32 by ODD test. The specificity of antibody of ASBg14 and ASBg32 was very high; however,the specificity of ASBg26 and ASBg04 was not very satisfying. The results showed that all the strains of B. glumae tested produced 500 bp specific fragments by the ISE and the direct PCR method,while others showed negative PCR result. Comparing the sensitivity of these 2 detecting methods, about 1 × 105cfu·mL- 1of the bacterial suspension were detected by direct PCR and below 1 × 103cfu·mL- 1can only be detected by immunocapture PCR method. In the detecting experiments of the artificial inoculated seeds,at least 2 seeds should be required by direct PCR; however,only 1 seed was enough for detection by immunocapture PCR.