Fluorescent quantitative PCR detection reagent for bacterial type I integron

Abstract

The invention discloses a fluorescent quantitative PCR detection reagent for a bacterial type I integron. The PCR technology is combined with fluorescence detection, a pair of primers and a probe with high amplification efficiency and high specificity are constructed and screened, the defects that the conventional PCR consumes time, and is easy to pollute and low in accuracy and electrophoresis detection is needed after amplification are overcome, the type I integron in bacteria can be rapidly, accurately, safely and qualitatively detected, the reagent has the advantages of simplicity, easiness in operation, intuitive result, high sensitivity and high repeatability, and the pollution risk of DNA or PCR products can be reduced. Moreover, the conventional PCR detection time is generally over 6 hours, the electrophoretic analysis after amplification is not needed, the sample detection time is within 3 hours, the positive, negative and uncertain results can be judged, the Ct value is 40.0 and can be determined as the positive or negative critical value, a plurality of samples can be detected, and the reagent has the high flux property.

Description

technical field [0001] The invention relates to PCR detection of integrons, in particular to a fluorescent quantitative PCR detection reagent for bacterial type I integrons. Background technique [0002] Integron is a site-specific genetic recombination system that can recognize and capture mobile gene cassettes. The increasing pressure of antibiotic application has led to the emergence of more and more drug resistance gene cassettes, and integrons carrying recombinant gene cassettes can be randomly inserted into conjugative plasmids or transposons, resulting in a wider range of horizontal transmission of drug resistance . The dissemination of integrons by means of transformation, transduction, and conjugation not only occurs between bacteria, but also across the boundaries of fungi and bacteria, and there is evidence that bacterial integrons are also increasing year by year in bacteria trend. Therefore, the rapid detection of integrons in the laboratory is similar to th...

AI Technical Summary

Problems solved by technology

For example, the invention patent with the authorized announcement number CN101948909 discloses the PCR method for detecting bacterial type I, II, and III integrons. The PCR detection of type I integrons has good sensitivity and specificity, but DNA electrophoresis is required. PCR post-processing, the detection time is long, and the toxic reagent ethidium bromide is exposed at the same time, the safety is poor, cross-contamination is prone to occur, resulting in false positive results, and the accuracy of the detection results is poor