Rapid detection method of blood pathogenic bacteria based on CE-SSCP (Capillary Electrophoresis-Single Strand Conformation Polymorphism)

Abstract

The invention discloses a rapid detection method of blood pathogenic bacteria based on CE-SSCP (Capillary Electrophoresis-Single Strand Conformation Polymorphism). In the rapid detection method, specific to conserved regions of 16SrRNA genes of staphylococcus aureus, escherichia coli, pseudomonas aeruginosa, staphylococcus epidermidis, klebsiella pneumoniae, enterococcus faecalis and streptococcus pneumoniae which are infected by blood, a corresponding PCR (Polymerase Chain Reaction) general primer is designed and subjected to fluorescence labeling, and then PCR is adopted to carry out amplification and electrophoresis detection on pathogenic bacteria in a blood sample, wherein the length of the designed general primer is suitable to the span of the amplification and has high complementary degree with the DNA of a pathogenic bacteria template in the blood sample to be tested, which enables an amplification product to have good specificity and favorable suitability to same kind of bacterium of different bacterial strains. The method has good repeatability and high sensitivity, can detect trace amount of the DNA of a pathogenic bacteria genome and is very suitable to wide application clinically.

Description

technical field [0001] The invention relates to a detection method for pathogenic bacteria infected with blood, in particular to a CE-SSCP-based rapid detection method for pathogenic bacteria in blood. Background technique [0002] Blood infection often occurs in high-risk inpatients who are immunocompromised or undergoing invasive procedures. After infection, the symptoms of poisoning are severe and the mortality rate is high, which is very difficult in clinical work. Traditional detection methods for blood infection pathogenic bacteria mainly rely on biochemical identification after direct culture. This method requires separate cultivation and identification of each pathogen, which generally takes 3-5 days, which is time-consuming, and the test results are greatly affected by factors such as blood collection dose, time of inspection, and timing of medication. [0003] Capillary electrophoresis (CE) is a new and efficient analysis and separation technology developed from t...

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Problems solved by technology

[0004] Today, CE-SSCP (capillary electrophoresis-single-strand conformation polymorphism) technology is widely used in the fields of gene point mutation screening, microbial classification, new species identification and molecular epidemiology, and is used for gene point mutation screening and common pathogen detection It has been reported, but the repeatability and reliability of this CE-SSCP technology largely depend on the selection and setting of various indicators in the operation process, such as the design of primers, the electrophoresis voltage intensity, etc.

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