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Multiplex PCR detection kit for common bacterial pathogens in cattle

A kit and a technology for bacterial diseases, which are applied in the field of detection kits for detecting Brucella bovis, Mycobacterium bovis and Chlamydia abortus, can solve problems such as no disclosure, avoid wasting materials, have good clinical application prospects, Avoid the cumbersome effect of the detection process

Active Publication Date: 2020-01-07
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • Multiplex PCR detection kit for common bacterial pathogens in cattle
  • Multiplex PCR detection kit for common bacterial pathogens in cattle
  • Multiplex PCR detection kit for common bacterial pathogens 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

The invention relates to a detection kit for detecting and distinguishing single or combined infection with three kinds of common pathogens in cattle by designing three pairs of specific amplificationprimers for three kinds of pathogens based on brucella abortus, mycobacterium bovis and chlamydophila abortus genes and cooperating with a PCR reaction component to establish a multiplex PCR method,or for detecting and distinguishing brucella abortus, mycobacterium bovis and chlamydophila abortus after packaging.

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...

Claims

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Application Information

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Patent Type & Authority Applications(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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