Quantitative polymerase chain reaction (qPCR) detection method for residual viable bacteria and spores of production strain in vitamin B2 as well as primers and probes used in qPCR detection method

Abstract

The invention relates to the technical field of biology, in particular to a quantitative polymerase chain reaction (qPCR) detection method for residual viable bacteria and spores of a production strain in vitamin B2 as well as primers and probes used in the qPCR detection method. The qPCR detection primers for the residual viable bacteria and spores of the production strain in vitamin B2 are designed. The production strain is B.subtilius KCCM-10445. The nucleotide sequences of the primers are as follows: the forward primer is 5'-CGA GCT TTT GCG CGT ATA-3', and the reverse primer is 5'-GCC ATTCCA ATA CAA AAC CAC ATA-3'. The probes, a kit and the detection method are also designed. The residual viable bacteria and spores of the production strain B.subtilius KCCM-10445 in a vitamin B2 product can be accurately detected, and the primers, the probes and the detection method related to detection are developed.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to vitamin B 2 The qPCR detection method and the primers and probes used for the residual viable bacteria and spores of the production strains in medium. Background technique [0002] Vitamin B 2 Also known as riboflavin, it is slightly soluble in water and is stable when heated in neutral or acidic solution. It is an integral part of the prosthetic group of yellow enzymes in the body (yellow enzyme plays the role of hydrogen delivery in biological redox). When it is lacking, it will affect the biological oxidation of the body, cause metabolic disorders, and lead to various diseases. It is indispensable for life activities. vitamins. For humans and animals, vitamin B must be obtained from food 2 . The human body needs about 0.3-1.8mg vitamin B per day 2 , while animal feed must contain 1-4mg / kg vitamin B 2 In order to meet its growth needs and improve nutrient utilization. [00...

AI Technical Summary

Problems solved by technology

The 16S rRNA gene sequence analysis method is commonly used to identify bacteria, but this method is cumbersome and has limitations in the identification of transgenic strains, and the 16S rRNA gene similarity of Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus licheniformis reaches more than 99%. This method cannot be used to distinguish, so we use the unique sequence of the GMB.subtilis strain to design qPCR primers and probes, which can specifically identify the residues of the production strain

Images