Amplification primer group of pseudomonas aeruginosa, applications of amplification primer group, and detection method of pseudomonas aeruginosa

Abstract

The invention provides an amplification primer group of pseudomonas aeruginosa, applications of the amplification primer group, and a detection method of pseudomonas aeruginosa. The detection method of pseudomonas aeruginosa comprises the following steps: (1) extracting the genome of a pseudomonas aeruginosa sample to be detected; (2) providing the specific gene oprL amplification primer group ofpseudomonas aeruginosa: CP1, CP2, F1, F2, D1, C1, R1, D2, C2, R2, C1*, C2* as well as D1* and D2*; (3) in a multi-crossing isothermal amplification reaction system comprising the amplification primergroup, carrying out isothermal amplification by adopting the genome nucleic acid of the pseudomonas aeruginosa sample to be detected as the template, thus obtaining the amplification product; and (4)detecting the amplification product of the step (3) by adopting a gold nanobiosensor. For the method provided by the invention, the multi-crossing isothermal amplification is adopted as the basis, thenanometer biological detection technology is combined, and pseudomonas aeruginosa can be rapidly and sensitively identified with high specificity.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to an amplification primer set for Pseudomonas aeruginosa, its application and a detection method for Pseudomonas aeruginosa. Background technique [0002] Pseudomonas aeruginosa (PA), also known as Pseudomonas aeruginosa, belongs to the genus Pseudomonas, is an aerobic Gram-negative bacillus, and is also a common opportunistic pathogen. The bacteria exist in soil, dust, water and a small number of human intestines, and can also temporarily parasitize the skin, mainly in the anus, genitals, armpits and external auditory canal. Typical strains produce two pigments, blue-green pyocyanin and yellow-green pyocyanin. Under normal circumstances, Pseudomonas aeruginosa does not cause disease. die. Systemic Pseudomonas aeruginosa infection can lead to sepsis. Patients have fever, jaundice, splenomegaly, and pneumonia, urinary tract infection, and meningitis can oc...

AI Technical Summary

Problems solved by technology

At present, clinical isolation and identification of Pseudomonas aeruginosa mainly relies on traditional bacterial culture and biochemical identification. This method includes secondary enrichment, selective culture and subsequent biochemical identification, which takes at least 3 days. Although the results of biochemical identification can be Relying on the interpretation of the automatic microbial identification instrument, but the disadvantage of this method is time-consuming and labor-intensive, which seriously delays the time of clinical diagnosis and treatment

With the rapid development of nucleic acid diagnostic technology, some PCR-based diagnostic technologies (such as ordinary PCR technology, fluorescent PCR technology) have been used for the rapid diagnosis of Pseudomonas aeruginosa, but these methods rely on expensive and complicated equipment and Expensive probe synthesis with subsequent electrophoresis steps and skilled operators

Some grassroots laboratories cannot meet the above conditions, thus limiting the application of these technologies at the grassroots level

In addition, the current PCR method and real-time PCR method to detect Pseudomonas aeruginosa, the detection process also takes 2-4 hours, which is not conducive to rapid detection and emergency detection

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