The invention discloses an RPA (
recombinase polymerase amplification) primer composition and method for detecting
shigella toxin-producing
Escherichia coli. The primer composition is used for detecting nine
gene subtypes, namely, stx1a, stx1c, stx1d, stx2a, stx2b, stx2c, stx2d, stx2e and stx2g and comprises a primer pair with sequences shown as SEQ ID NO: 1 and SEQ ID NO: 4 and a primer pair withsequences shown as SEQ ID NO: 9 and SEQ ID NO: 12. The method provided by the invention has the advantages of simple operation, high specificity and high sensitivity, is suitable for on-site
rapid detection, can complete
nucleic acid detection within 20 min, and can provide a rapid
screening result for supervision and inspection of listed products and
emergency treatment of supervision departments. Meanwhile, the primer and probe combination designed and screened for the first time can effectively detect nine stx
gene subtypes, namely stx1a, stx1c, stx1d, stx2a, stx2b, stx2c, stx2d, stx2e andstx2g genes. And in combination with a centrifugal disc type micro-fluidic
chip technology, rapid and high-
throughput detection on STEC in foods, medicines and
livestock breeding products can be realized within an extremely small volume (5 microliters ).