Primers, kit and method for detecting Escherichia coli Shiga toxin I by PSR (polymerase spiral reaction)

Abstract

The invention discloses primers, kit and method for detecting Escherichia coli Shiga toxin I by PSR (polymerase spiral reaction). The primers for detecting Escherichia coli Shiga toxin I are designedaccording to specific target sequence stx1 of Shiga toxin I and include detection primer Ft and detection primer Bt, as well as acceleration primer IF and acceleration primer IB, and their nucleotidesequences are shown as SEQ ID NO. 1 to 4. The invention also provides a kit for detecting Escherichia coli Shiga toxin I by PSR; the kit includes the above primers, Bst DNA polymerase, and a mixed solution of calcein and manganese chloride, is suitable for detecting Escherichia coli Shiga toxin I by polymerase spiral reaction; after developing with a fluorescent dye, the results can be judged witheyes. The kit is simple and quick to operate, low in detection cost and suitable for field detection.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a primer, a kit and a method for detecting Escherichia coli type I Shiga toxin by PSR. Background technique [0002] Shiga toxin is mainly produced by Escherichia coli, and Shiga toxin-producing Escheriachia coli (STEC) is the main pathogenic factor of human food poisoning in the world, and it is also the main cause of large-scale foodborne food poisoning. pathogenic bacteria. There are more than 100 serotypes of pathogenic Escherichia coli in STEC that can cause disease, and can cause non-hemorrhagic diarrhea, hemorrhagic colitis, hemolytic uremic syndrome and other diseases. The pathogenic characteristic virulence factors are Shiga toxin type I (stx1) and Shiga toxin type II (stx2), encoded by stx1 and stx2 respectively, and diseases caused by Stx1 are often reported. [0003] Currently, Shiga toxin detection methods mainly include conventional detection methods, PCR detection me...

AI Technical Summary

Problems solved by technology

Immunological detection methods such as ELISA method are to adsorb immunologically active antigens or antibodies on the surface of a solid phase carrier, add an active enzyme-labeled antibody or antigen, and then add an enzyme chromogenic substrate, and the active enzyme acts on the substrate. Judging by producing color, this method has high sensitivity and strong specificity, but its operation process is complicated, there are many influencing factors, and the cost is high, so it is not suitable for mass detection

The gene chip detection method developed in recent years has the characteristics of high throughput, rapidity, and high degree of automation. However, due to the high cost of this technology, it cannot be widely used in the field of routine detection.

Although the most widely used constant temperature amplification technology-LAMP has the advantages of high sensitivity and specificity, it also has disadvantages such as complex primer design, high false positive rate, and high reagent prices.

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