Constant-temperature detection method for identifying lactobacillus brevis as well as special primer and kit thereof

Abstract

The invention discloses a constant-temperature detection method for identifying lactobacillus brevis as well as a special primer and a kit thereof. The specific primer group provided by the inventionis composed of a primer F3, a primer B3, a primer FIP, a primer BIP, a primer LF and a primer LB, and the six primers are sequentially shown as sequences 2-7 in a sequence table. The application of the specific primer group is as follows (b1) or (b2): (b1) identifying or assisting in identifying lactobacillus brevis; (b2) detecting whether the to-be-detected sample contains lactobacillus brevis ornot. The specific gene segment of lactobacillus brevis is amplified by using a loop-mediated isothermal amplification technology, the blank of the LAMP detection method of lactobacillus brevis in beer is filled, and the detection method has the advantages of strong specificity, simplicity in operation, rapidness, convenience and the like, can be used for beer quality monitoring, and is suitable for basic application and on-site rapid detection of winery.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a constant temperature detection method for identifying Lactobacillus brevis, as well as special primers and a kit. Background technique [0002] Lactobacillus breris is widely distributed in nature, belongs to Lactobacillus order, Lactobacillus family, and Lactobacillus genus, and is commonly found in milk, yoghurt, cheese, and sour pickles. When Lactobacillus brevis exists in beer, it often causes the acidity and viscosity of beer to increase, and what's more, it causes beer turbidity and precipitation, which affects the taste and quality of beer. Beer quality incidents caused by Lactobacillus brevis contamination are not uncommon. According to statistics, more than 50% of daily beer contamination incidents are caused by Lactobacillus brevis. Rapid detection of Lactobacillus brevis has become an urgent problem to be solved in the quality control of breweries. [0003] At present, th...

AI Technical Summary

Problems solved by technology

Among all current detection methods, this method has high accuracy, low cost, and low requirements for experimenters, and can meet the needs of grassroots applications. However, no matter which medium is used for culture and identification, the detection of the results requires 5 More than 7 days, unable to reflect the beer pollution in time, unable to meet the needs of rapid quality monitoring

Bioluminescence and immunology methods mainly include ATP bioluminescence detection technology and antigen-antibody detection method; ATP bioluminescence detection technology achieves the purpose of detecting microorganisms by detecting ATP produced in the metabolic process of microorganisms, and the detection results can be obtained quickly when using this method for detection , can also quantify microorganisms, but cannot qualitatively identify different microorganisms, and is susceptible to external interference, resulting in unstable results; the antigen-antibody method is based on the principle of specific combination of antigen and corresponding antibody to form a precipitation complex for qualitative detection of microorganisms, including agglutination reaction, precipitation reaction, ELISA, etc. This microbial diagnostic method has the advantages of strong specificity, but the antibody screening process is complicated, the cost of reagent detection is expensive, and it is easily affected by human interference, which cannot meet the detection needs of wineries at the grassroots level

[0004] Molecular biology methods are based on nucleic acid level detection. Common molecular biology methods applied to microbial detection include fluorescence in situ hybridization (FISH), gene chip technology, and polymerase chain reaction PCR. The advantage of these methods is specificity. Strong and time-consuming, but most of them are difficult to be effectively carried out in winery testing due to complex operations and high requirements for equipment and operators.

Images