Multiplex pcr detection kit for pathogens of common bacterial diseases in cattle

A kit and technology for bacterial diseases, applied in the field of kits for common bacterial disease pathogens in cattle, can solve the problems of undisclosed and other issues, and achieve the effects of avoiding waste of materials, good clinical application prospects, and avoiding cumbersome detection processes

Active Publication Date: 2022-03-25
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
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  • Multiplex pcr detection kit for pathogens of common bacterial diseases in cattle
  • Multiplex pcr detection kit for pathogens of common bacterial diseases in cattle
  • Multiplex pcr detection kit for pathogens of common bacterial diseases in cattle

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[0017] In this example, cow feces were selected for detection of Brucella, Mycobacterium and Chlamydia abortus infection. Take an appropriate amount of cow feces, inactivate the bacteria at 70°C for 30 minutes, use the fecal DNA extraction kit to extract the whole genome DNA, and measure the concentration of the extracted DNA for later use. Prepare a multiple PCR reaction solution in a PCR reaction tube, using a 25 μL reaction system, including 5 μL of 5×PCRBuffer, 12.5 μL of 2×Taq DNA polymerase Mix, and 100 ng of cow dung whole genome DNA template for Brucella The upstream primer F1 for specific detection: 5'-CACTCGTGCTAGCAATTTTCTCGC-3' (SEQ ID №1). 0.5 μL (10 μmol / L) and downstream primer R1: 5'-CGACATTGACCGATACGTTATAGC-3' (SEQ ID №2) 0.5 μL (10 μmol / L), upstream primer F2 for specific detection of M. bovis: 5' - GGCCTGGGCGGCACTGTG-3' (SEQ ID №3) 0.5 μL (10 μmol / L) and downstream primer R2: 5'-CAGACTGTCGTCGGTGATAGC-3' (SEQ ID №4) 0.5 μL (10 μmol / L) for Chlamydia abortus S...

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Abstract

Based on the genes of Brucella abortus, Mycobacterium bovis and Chlamydia abortus, the present invention designs three pairs of specific amplification primers for the three pathogens, cooperates with the PCR reaction components, and establishes a multiplex PCR method for detecting and distinguishing the three kinds of cattle Single or mixed infection of common pathogens, and packaging of detection kits for the detection and differentiation of Brucella bovis, Mycobacterium bovis and Chlamydia bovis.

Description

technical field [0001] The invention relates to a kit for common bacterial disease pathogens in cattle, in particular to a kit for detecting Brucella bovis, Mycobacterium bovis and Chlamydia bovis abortus. Background technique [0002] Brucella abortus ( Brucella abortus ), mycobacteria ( Mycobacterium bovis ) and Chlamydia abortus ( Chlamydia abortus ), is the most common bacterial disease pathogen in cattle, among which Brucella can cause abortion, orchitis, tenosynovitis and arthritis in cattle; Mycobacterium bovis can cause nodules in various tissues and organs such as lungs, lymph nodes and colon Chlamydia bovis can cause abortion in cows, stillbirths, weak fetuses, endometritis, orchitis in bulls, enteritis in calves, conjunctivitis, etc. symptom. These three pathogens are also chronic zoonotic pathogens, which can cause similar clinical symptoms and damage to tissues and organs in the human body, pose a threat to human health and even life safety, and cause inca...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04
CPCC12Q1/689C12Q1/686C12Q2600/16C12Q2537/143
Inventor 娄忠子周继章李兆才刘星谭书敏
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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