Application of DetectingVibrio Cholerae Combined with Serum Agglutination and Gene Sequencing / 现代检验医学杂志

ABSTRACT

Objective To avoid false positive detection ofVibrio cholerae and improve the detection correct rate.Methods 1~7 months of 2013 were randomly selected,the national various provinces and cities CDC to China cholera CDC positive screening 14 strains.LB nutrient agar 12 hours,take single colony to Vibrio cholera serum agglutination,extraction of strain DNA at the same time boiled template method.For Vibrio 16SrDNA sequence and design primers for PCR detection of Vib-rio,16SrDNA,electrophoresis were used to observe the 16SrDNA products,16SrDNA positive products sent to sequencing company sequencing,sequencing results were Blast comparison on the NCBI website for the analysis and comparison of ser-um agglutination and Blast alignment.Results 12 strains was positive for agglutination and 2 strains of non agglutination in 14 strains.The Vibrio 16SrDNA amplification,electrophoresis were used to observe the 14 isolates that were amplified frag-ment corresponding,that the selected strains were vibrio.The 16SrDNA positive products were 14 strains,and the sequen-cing the Blast results:2 strains of bacteria were Vibrio harveyi for non agglutination,in 12 positive strains of serum aggluti-nation;1 strains was Vibrio natriegen and 11 strains wereVibrio cholerae .Conclusion Detection ofVibrio cholerae cholerae combined with serum agglutination and gene sequencing can avoid false positive result ofVibrio cholerae ,and improve cor-rect rate of the detection.