Comparative study of different methods for detecting foodborne pathogens / 国际检验医学杂志

ABSTRACT

Objective To investigate the feasibility of the gene chip technique and multiplex PCR technique for detecting and screening foodborne pathogens .Methods The primer sequences were designed to target cells ,the biotin was adopted to label the re‐verse primer 5′end and the amino group was adopted to label oligonucleotide probe 5′end .The probe spotted on a solid support for preparing microarray ,PCR product was hybridized with microarray probe region ,PCR and hybridization reaction system was opti‐mized .Results The microarray technique could simultaneously detect multiple pathogens of Shigella ,Salmonella ,Klebsiella pneu‐moniae ,Brucella ,Proteus mirabilis ,Staphylococcus aureus ,Campylobacter jejuni ,etc .,which was easy to operate and had strong specificity .The sensitivity of bacterial pure cultures was 5 .0 × 102 CFU/mL ,the sensitivity of DNA detection was 0 .1 pg ,the coin‐cidence rate for detecting isolated bacteria was 100 % .The PCR detection system was established and optimized by using primers , the concentration of Mg2+ and the annealing temperature Tm value of 1 .5 mmol/L and 56 ℃ were determined ,the detection sensi‐tivity reached to 10 pg ,all the specific primers amplified bands could be amplified under this sensitivity .Conclusion The gene chip technique and multiplex PCR technique can effectively detect foodborne pathogens ,which provide a new idea for detecting pathogens with the high‐throughput screening and are worth popularization and application in the field of food safety .