Establishment and application of a rapid PCR detection method of Brucella spp / 国际检验医学杂志

ABSTRACT

Objective To establish a polymerase chain reaction (PCR) method for detecting Brucella spp .and to evaluate its ap‐plication .Methods The PCR primer aiming at the outer membrane protein(omp‐2) coding gene were designed and the PCR method for detecting Brucella spp .was established .By using DNA of 19 kinds of common bacteria and Brucella spp .,the specificity and sensitivity were evaluated .Three strains of suspected Brucella spp .were performed the nucleic acid detection .Then ,3 suspected strains of Brucella spp .were amplified by the PCR method with omp‐2 gene ,and the results were compare to the those by the cul‐ture method .Results The established Brucella spp .PCR detection method could only detect the Brucella spp .positive strain ,the control bacterium DNA did not appear the target band ;the sensitivity was 100 copies/response;the PCR method detected omp‐2 gene band in 3 strains of suspected Brucella spp .,which was identified as Brucella spp .and was same to the results by the isolation culture method .Conclusion The established PCR method for detecting Brucella spp .is accurate and rapid compare with the isola‐tion and culture method ,which is suitable for the rapid detection of brucellosis epidemic situation .