Detection kit based on hybridization chain reaction (HCR) fish pathogenic microorganism aeromonas hydrophila metG gene and application thereof

• Main Authors: CHEN ZHILAN, SUN YUANDONG, ZHONG LIZHEN, DENG JIMIN, ZHANG YIFEI, CUI XIAOJUAN
• Format: Patent
• Language: Chinese; English
• Published: 27.02.2024
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention relates to development and application of a detection kit based on a hybridization chain reaction (HCR) fish pathogenic microorganism aeromonas hydrophila metG gene. The detection kit comprises a detected aeromonas hydrophila specific DNA sequence metyl-tRNA synthase, a pair of hairpin primers metG-H1 and metG-H2, a fluorophore, a quenching group, a DNA storage solution (1X) and a CaCl2 solution. When in use, the H1 and H2 reagents are added into a water body sample or a bacterial liquid prepared by separating visceral organs and tissues of sick fishes; standing on ice, and carrying out heat shock at a constant temperature of 42 DEG C; quickly placing on ice for cooling after heat shock is completed; and adding a DNA storage solution, placing at room temperature, and detecting by using a fluorescence microscope. And judging whether the sample to be detected contains aeromonas hydrophila or not and the proportion of pathogenic bacteria by observing the luminescence condition of the sample under a