DNA biosensor for rapidly detecting clostridium perfringens in meat products and detection method thereof

A clostridium perfringens and biosensor technology, applied in the field of detection, can solve the problems of easy occurrence of false positives, high detection cost, long-term verification, etc., and achieve the effects of low cost, high sensitivity, and improved accuracy

Pending Publication Date: 2022-08-09
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Traditional microbial detection methods require long-term anaerobic culture and complicated verification, and have disadvantages such as time-consuming and high detection limits; although immunological methods can achieve rapid detection through antigen-antibody binding reactions, they are prone to false positives; biomolecular detection methods Has high sensitivity, but its detection cost is high

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  • DNA biosensor for rapidly detecting clostridium perfringens in meat products and detection method thereof
  • DNA biosensor for rapidly detecting clostridium perfringens in meat products and detection method thereof
  • DNA biosensor for rapidly detecting clostridium perfringens in meat products and detection method thereof

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Embodiment 1

[0044] Example 1: Detection of Clostridium perfringens

[0045] 1.1 Screening of Clostridium perfringens-specific primers

[0046] Methods: Pseudomonas putida (CICC 20677), Klebsiella pneumoniae (CICC 20093) and Escherichia coli (CICC C0031) were extracted from TIANamp Bacteria DNA Kit (Tiangen Biochemical Technology Co., Ltd., Beijing). , Pseudomonas aeruginosa (CICC 21625), Lactobacillus plantarum (CICC 6240), Bacillus subtilis (CICC25064), Salmonella (CICC B0082), Staphylococcus aureus (CICC B0172), Clostridium sp. (ATCC 17787), The genome of Clostridium perfringens (CICC 22949) was amplified by PCR with primers CPA-F (5'-GCTAATGTTACTGCCGTTGA-3') and CPA-R (5'-CCTCTGATACATCGTGTAAG-3') using the extracted genome as a template. The PCR amplification procedure was pre-denaturation at 94 °C for 4 min; denaturation at 94 °C for 40 s, annealing at 52 °C for 1 min, extension at 72 °C for 30 s, 35 cycles; extension at 72 °C for 10 min, PCR products were verified by agarose gel ele...

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Abstract

The invention discloses a DNA (deoxyribonucleic acid) biosensor for rapidly detecting clostridium perfringens in meat products and a detection method of the DNA biosensor. The biosensor comprises nano magnetic beads, DNA walker, DNA Aptamer, DNA H1, DNA H2, DNA Trigger, DNA H3 and DNA H4, wherein carboxyl groups are modified on the surfaces of the nano magnetic beads, and the nucleotide sequences of the DNA walker, the DNA Aptamer, the DNA H1, the DNA H2, the DNA Trigger, the DNA H3 and the DNA H4 are respectively shown as SEQ ID NO: 1-7. By introducing the magnetic nanoparticles and orderly assembling the nucleic acid aptamers on the surfaces of the magnetic nanoparticles, the operation steps in the detection process of the clostridium perfringens are greatly simplified, and the detection cost of the clostridium perfringens is reduced. Meanwhile, the detection method provided by the invention has the advantages of high sensitivity, low cost, rapidness and the like.

Description

technical field [0001] The invention relates to the field of detection, in particular to a DNA biosensor for rapidly detecting Clostridium perfringens in meat products and a detection method thereof. Background technique [0002] Clostridium perfringens is a Gram-positive anaerobic bacterium widely distributed in the environment, C. perfringens can grow in a wide temperature range of 5-50 °C, and 37-45 °C is its optimum temperature. At high growth temperatures, spores can survive for several years under harsh environmental conditions. C. perfringens can produce toxins that can easily lead to food-borne illnesses, including diarrhea and severe abdominal pain, and is a zoonotic pathogen. At present, the methods of detecting Clostridium perfringens by various means such as microbiology, immunology and biomolecular science are gradually being developed at home and abroad. [0003] Traditional microbial detection methods require long-term anaerobic culture and complicated verif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/327G01N21/64
CPCG01N27/26G01N27/3278G01N21/6428Y02A50/30
Inventor 孙芝兰刘芳王文卓王德宝王道营诸永志徐为民
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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