Amplification primer sets of Klebsiella pneumoniae, application and method for detecting Klebsiella pneumoniae

Abstract

The invention provides amplification primer sets of Klebsiella pneumoniae, application and a method for detecting the Klebsiella pneumoniae. The method for detecting the Klebsiella pneumoniae comprises the following steps: 1) extracting a genome of a Klebsiella pneumoniae sample to be detected; 2) providing the following specific gene rcsA amplification primer sets of the Klebsiella pneumoniae: CP1, CP2, F1, F2, D1, C1, R1, D2, C2, R2, C1*, C2*, D1* and D2*; 3) using genomic nucleic acid of the Klebsiella pneumoniae sample to be detected as a template for isothermal amplification under a multi-crossover isothermal amplification reaction system comprising the amplification primer sets to obtain an amplification product; 4) detecting the amplification product of the step 3) by using a gold nanobiosensor. The method provided by the invention uses multi-crossover isothermal amplification as a basis, is combined with a nanometer biological detection technology, and can rapidly, sensitivelyand highly specifically identify the Klebsiella pneumoniae.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to an amplification primer set for Klebsiella pneumoniae, an application thereof and a detection method for Klebsiella pneumoniae. Background technique [0002] Klebsiella pneumoniae (Klebsiela Pneumoniae, K.P), commonly known as Klebsiella pneumoniae, is one of the most important pathogenic bacteria in clinical isolation and hospital infection. Disease-causing accounts for more than 95% of Klebsiella infections. Klebsiella pneumoniae is a gram-negative bacillus that exists in the upper respiratory tract and intestinal tract of the human body. When the body's resistance is reduced, it enters the lungs through the respiratory tract and causes confluent consolidation of the lobules or lobules, and the exudate in the lesion is viscous. Thick and heavy, causing the leaf gap to drop. In nosocomial sepsis, Klebsiella, Pseudomonas aeruginosa, and Serratia are imp...

AI Technical Summary

Problems solved by technology

Rapid diagnosis of pathogenic bacteria is the cornerstone of effective treatment of such diseases, but the current clinical isolation and identification of Klebsiella pneumoniae mainly relies on traditional bacterial culture and biochemical identification, which includes secondary enrichment, selective culture and subsequent biochemical identification , it takes at least 3 days. Although the results of biochemical identification can be interpreted by the automatic microbial identification instrument, 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 Klebsiella pneumoniae, 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 Klebsiella pneumoniae detection process also takes 2-4 hours, which is not conducive to rapid detection and emergency detection

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