Loop-mediated isothermal amplification primers, kits and methods for detecting 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 for detecting Klebsiella pneumoniae and a detection method. Background technique [0002] Klebsiella pneumoniae is a gram-negative, non-exercise, capsular, lactic acid fermentation, and facultative anaerobic rod-shaped bacteria. Klebsiella pneumoniae usually resides in the intestines of rats, mice and other animals. As an opportunistic pathogen, once Klebsiella pneumoniae enters the respiratory tract, it can infect rats and mice, causing severe lung damage, and the bacteria has a stronger growth tendency to replace other bacteria in antibiotic-resistant nude mice. One of the common health monitoring indicators for rodents. [0003] The currently commonly used methods for detecting Klebsiella pneumoniae include the direct plating method using monitoring swabs, the chromogenic medium method and the conventional PCR method / fluorescence quantitative PCR...

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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