Loop-mediated isothermal amplification (LAMP) primers, kit and method for detecting mouse Klebsiella pneumoniae

Abstract

The invention discloses loop-mediated isothermal amplification (LAMP) primers, a kit and a method for detecting mouse Klebsiella pneumoniae. LAMP primers are designed by using the Klebsiella pneumoniae tyrB gene sequence to establish a high-specificity high-sensitivity Klebsiella pneumoniae visual detection method suitable for clinical sample detection. The green fluorescence is observed to quickly and accurately detect the existence of Klebsiella pneumoniae, wherein the lowest detection limit is 1.6 number of copies / reaction. The 6 designed and screened primers are used for identifying the specific sequence region of the target sequence, thereby ensuring the high specificity of LAMP amplification. The added loop primers greatly enhance the efficiency, have the advantage of short consumption time and are suitable for field application of the 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, immobile, 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-consuming, medium, operation steps or culture conditions are quite different in different laboratories, so the results may be inconsistent or false negative / positive

The conventional PCR method and the 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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