Microdroplet digital PCR (Polymerase Chain Reaction) detection method for enterococcus hisei in medical food

• Main Authors: ZHANG JIE, TANG XIAOJUAN, LIU YUNGUO, LIU KE, LEI ZHIWEN
• Format: Patent
• Language: Chinese; English
• Published: 20.07.2021
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROCESSES USING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a PCR (Polymerase Chain Reaction) amplification primer and a probe primer of Enterococcus hirsutei. The primer comprises the following components: a forward sequence of an enterococcus hirsutei primer: 5'-CCACTCAGCTTGATGTAA-3', a reverse sequence of an enterococcus hirsutei primer, a reverse sequence of an enterococcus A reverse sequence: 5'-GTTTGTTCAATAATCTTCAGATC-3', the sequence of the probe is 5'-6-FAM-ACAACAGCACCACGACGAT-TAMRA-3'. A microdroplet digital PCR detection method for enterococcus hisei in medical food comprises the following steps: weighing 25g of a sample, putting the sample into a homogenizing bag filled with 225mL of sterile water, fully and uniformly mixing, and diluting in a 10-fold gradient manner to obtain a sample to be detected; extracting DNAs of a to-be-detected sample, a positive control and a negative control; acquiring a PCR amplification primer and a probe for identifying the enterococcus hirsutei; and carrying out droplet digital PCR amplification on the