LAMP (loop-mediated isothermal amplification) primers, kit and method for detecting murine klebsiella pneumoniae

Abstract

The invention discloses LAMP (loop-mediated isothermal amplification) primers, a kit and a method for detecting murine klebsiella pneumoniae. A visual detection method of murine klebsiella pneumonia which is suitable for clinical sample detection and has strong specificity and high sensitivity is established by designing the LAMP primers by virtue of a klebsiella pneumoniae tyrB gene sequence. Whether klebsiella pneumoniae exists or not can be detected quickly and accurately by observing whether green fluorescence exists or not, and the lowest detectable limit is 5.6 copy number / reaction. According to identification of a specific sequence region of a target sequence by four primers designed and screened, high specificity of LAMP diffusion is guaranteed, so that the LAMP primers are suitable for field application at basic level.

Description

technical field [0001] The invention belongs to the technical field of medical microorganism detection, and relates to a LAMP primer and a detection method for detecting Klebsiella pneumoniae. Background technique [0002] Klebsiella pneumoniae is a Gram-negative, immotile, encapsulated, lactic acid fermenting, facultatively anaerobic, rod-shaped bacterium. Klebsiella pneumoniae commonly colonizes the guts of mice, mice and other animals. As an opportunistic pathogen, once Klebsiella pneumoniae enters the respiratory tract, it can infect mice and cause severe lung damage, and the bacterium has a stronger growth tendency to replace other bacteria in antibiotic-resistant nude mice. One of the common health monitoring indicators used in rodents. [0003] The currently commonly used detection methods for Klebsiella pneumoniae include the direct plating method using monitoring swabs, the chromogenic medium method, and the conventional PCR method / real-time quantitative PCR metho...

AI Technical Summary

Problems solved by technology

However, compared with other methods, the bacterial culture method has disadvantages such as difficulty in standardization, long time consumption, and large differences in culture medium, operation steps or culture conditions in different laboratories, so the results may be inconsistent or false negative / positive

Conventional PCR method and fluorescent quantitative PCR method need to go through denaturation, annealing, extension, and the whole process of DNA extraction takes about 2 to 4 hours to produce results, and require sophisticated and expensive instruments and reagents to quickly and easily carry out molecular unfavorable diagnosis

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