Positive control template for PCR detection on drug resistance gene, preparation method and kit thereof

Abstract

The invention discloses a positive control template for PCR detection on a drug resistance gene, a preparation method and a kit thereof, wherein the drug resistance gene is NDM-1, and the sequence of the NDM-1 gene is the sequence as shown in SEQ ID NO: 1. The method for preparing the positive control template comprises the following steps: a) carrying out point mutation on the NDM-1 gene sequence so as to obtain a mutant gene, wherein the sequence of the mutant gene comprises the sequence as shown in SEQ ID NO: 3; b) synthesizing the mutant gene and then cloning the mutant gene into plasmids; and c) linearizing the plasmids so as to obtain the positive template. By setting the positive template, the NDM-1 detection can be carried out quickly and accurately; and through carrying out the point mutation on the NDM-1 gene, that the positive template can be ensured not to be transcribed and translated correctly and only can be used as a sequence for positive control, thereby guaranteeing biosafety.

Description

technical field [0001] The invention relates to a positive control template for PCR detection of drug-resistant genes, a preparation method and a kit. Background technique [0002] With the widespread use of antibiotics, the problem of bacterial resistance is becoming more and more serious. In August 2010, The Lancet Infectious Diseases magazine published a joint research report by scientists from India, Britain, Sweden, Pakistan and Australia, reporting the prevalence of antibiotic resistance. A new mechanism and its spread in India, the United Kingdom and Pakistan, the problem of drug resistance has once again awakened the world that antibiotic misuse will end the power of our reliance on antibiotics to fight bacterial infections. [0003] As early as 1940, when penicillin was not yet clinically applied, scientists isolated the enzyme that hydrolyzes penicillin, which was called penicillinase at that time. After analyzing penicillin as an antibiotic with a β-lactam structu...

AI Technical Summary

Problems solved by technology

[0006] The easiest detection method is the polymerase chain reaction (Polymerase Chain Reaction, PCR) based on nucleic acid amplification in vitro. Although the NDM-1 gene sequence has been published online, it will make it difficult to judge the detection results without a positive control template. Moreover, If the published gene is directly synthesized, it will cause contamination of the synthetic gene to non-resistant strains

Images