Comparison of Three Detection Methods for Burkholderia Cepacia Complex
OBJECTIVE To establish a rapid and accurate PCR method for detecting 24 strains of Burkholderia cepacia complex(Bcc)by comparing three detection methods of loop-mediated isothermal amplification(LAMP),SYTO 9 dye method based on polymerase chain reaction(PCR)and TaqMan probe real-time fluorescent quantitative PCR method(TaqMan probe method).METHODS According to the molecular biological information of 24 strains of Bcc in the NCBI database,multiple candidate sequence fragments unique to Bcc were screened out,and specific primer and probe that could simultaneously detect 24 strains of Bcc were designed.At the same time,the detection methods of LAMP,SYTO 9 dye method based on PCR and Taqman probe were explored,and the optimal annealing temperature was optimized and screened.The 39 experimental strains were used to verify the Bcc detection method.RESULTS LAMP method could not effectively detect Bcc,SYTO 9 dye method and TaqMan probe method could effectively detect more than 20 strains of Bcc,while TaqMan probe method had higher amplification effect,better detection sensitivity,repeatability and stability,which could meet the requirements of this study.CONCLUSION In this study,a TaqMan probe method for rapid detection of Bcc was established.Compared with LAMP method and SYTO 9 dye method,this method has the advantages of fast,simple and high sensitivity,and provides technical support for the rapid detection of Bcc.
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