PCR (Polymerase Chain Reaction) detection kit for cat and/or dog pathogens, detection method and application
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A technology for detecting kits and pathogens, applied in the field of biomedicine, can solve the problems of low detection accuracy and low positive coincidence rate of pet pathogens, and achieve the effects of saving detection time, strong specificity, and saving human resources
Pending Publication Date: 2022-04-05
北京迈基诺基因科技股份有限公司
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[0004] For above-mentioned deficiencies in the prior art, the problem to be solved by the present invention is: how to provide a kind of PCR detection kit and detection method and application of cat and / or canine pathogen, to solve the detection of pet pathogens in the existing colloidal gold test paper technology. The problem of low precision and low positive coincidence rate
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Embodiment 1
[0141] Reagent test kit
[0142] 1. Specific primers and probes, see the table below
[0143]
[0144]
[0145]
[0146] 2. The PCR reaction reagent is: TaqPath TM 1-Step Multiplex Master Mix (No ROX).
[0147] 3. Deionized water.
Embodiment 2
[0149] DNA / RNA preparation
[0150] Collect 1ml of peripheral venous blood from cats and dogs according to routine medical procedures, and add EDTA for anticoagulation. Using Thermo's MagMAX TM CORE Nucleic Acid Purification Kit+MagMAX TM CORE Mastitis&PanbacteriaModule Extraction Kit, extract DNA and RNA.
Embodiment 3
[0152] PCR reaction system preparation
[0153] Prepare the PCR reaction system according to the following system:
[0154] A total of 8 reaction systems (the total reaction system is 20 μL), which are CS1-CS4 and FS1-FS4 in Table 1 and Table 2, respectively, were composed of mixed internal reference primers and probes.
[0155] PCR reaction reagent: 10 μL;
[0156] The concentration of each primer probe is 10M, the primer volume is 0.8 μL, and the probe volume is 0.4 μL;
[0157] Sample template: 2 μL;
[0158] Make up 20 μL with deionized water.
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Abstract
The invention relates to the technical field of molecular biomedicine, in particular to a PCR (Polymerase Chain Reaction) detection kit for cat and / or dog pathogens, a detection method and application. The kit is used for detecting canine distemper virus, canine influenza A virus, canine parainfluenza virus, canine parvovirus, canine coronavirus, canine rotavirus, canine babesia, canine ascaris, canine Ehrlichia, canine brucella, rabies virus, borrelia burgdorferi, reference gene ACTB, feline herpes virus, feline calicivirus and feline parvovirus. The kit comprises primers and probes of feline coronavirus, feline immunodeficiency virus, feline leukemia virus, feline mycoplasma, feline mycoplasma, feline chlamydia, giardia, toxoplasma, bartonella and reference gene GAPDH, collected DNA and RNA are added into the kit, a real-time fluorescence PCR instrument is adopted for PCR reaction, FAM, HEX, ROX and CY5 fluorescence signals are collected in each cycle, analysis of related pathogens is carried out, and the kit can be used for detecting the feline and the canine. Compared with a traditional detection method, the method has the advantages of higher specificity and higher sensitivity.
Description
technical field [0001] The invention relates to the technical field of biomedicine, in particular to a PCR detection kit, detection method and application of cat and / or dog pathogens. Background technique [0002] With the improvement of people's living standards, the number of pets in cities has soared. Pets have gradually become an important part of many families, and even treat each other as family members, and have very close contact with humans. However, while pets bring fun to people's lives, they also bring hidden dangers that are difficult to detect. threaten human health. The control of pet diseases has become one of the common concerns of the international community. Cats and dogs are currently the most common pets in China. According to incomplete statistics, there are about 80-400 million cats and dogs raised by humans around the world. Health issues are also a growing concern. [0003] At present, pet pathogen detection mainly includes colloidal gold test p...
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