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pcr-rflp primers and methods for identifying mycoplasma goat subspecies and other members of the mycoplasma cluster

A PCR-RFLP, mycoplasma technology, applied in the field of PCR-RFLP primers, achieves the effect of simple identification method and high accuracy

Active Publication Date: 2022-05-31
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

According to literature review, there is no PCR-RFLP method for Mycoplasma goat subspecies pgmb gene at home and abroad, but this method has been widely used in swine and poultry diseases

Method used

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  • pcr-rflp primers and methods for identifying mycoplasma goat subspecies and other members of the mycoplasma cluster
  • pcr-rflp primers and methods for identifying mycoplasma goat subspecies and other members of the mycoplasma cluster

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Embodiment 1

[0017] 1. Strain

[0018] Mycoplasma capricolum subspecies subspecies FJ-GT, the standard strain PG3 and Mycoplasma capricolum subspecies (GenBank accession number LN515398.1) isolated and identified in this laboratory were all herbivorous by the Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences Preserved by the Animal Disease Research Laboratory.

[0019] 2. Primer design and synthesis

[0020] The specific primers P1 and P2 were designed according to the differences in the restriction enzyme cleavage sites contained in the coding region of the pgmb protein gene of the Mycoplasma filamentosa subspecies and other members of the Mycoplasma filamentosa cluster. Downstream primer P2: 5'-GGTGTTATTTTTGATTTAGATGGAG-3'.

[0021] 3. PCR amplification

[0022] DNA was extracted using the instructions of Beijing Quanshijin Biotechnology Co., Ltd. DNA / RNA extraction kit, and the extracted DNA was used as a template for PCR amplification. P...

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Abstract

The present invention provides PCR-RFLP primers and methods for identifying Mycoplasma mycoplasma goat subspecies and other members of Mycoplasma mycoplasma cluster, the upstream primer P1: 5'-ACAAAAAATATTAGCATTCAAACTT-3', the downstream primer P2: 5'-GGTGTTATTTTTGATTTAG ATGGAG-3' , the method is to use the differences in the enzyme cutting sites contained in the pgmb protein gene coding region of mycoplasma goat subspecies and other members of the mycoplasma cluster to distinguish, including DNA extraction, PCR amplification to obtain the corresponding target fragment RFLP analysis was performed after HindⅢ digestion. The identification method of the invention has simple operation and high accuracy.

Description

technical field [0001] The invention belongs to the technical field of rapid diagnosis of veterinary infectious diseases, and in particular relates to a PCR-RFLP primer and method for identifying Mycoplasma filamentosa goat subspecies and other members of a Mycoplasma filamentosa cluster. Background technique [0002] Mycoplasma filamentosa goat subspecies (Mycoplasma mycoides subsp. capri, Mmc) is a member of the Mycoplasma filamentosa cluster, which is a Mycoplasma pneumonia of goats and sheep , MPGS) one of the pathogens. MPGS is highly contagious, mainly through respiratory tract infection, and can also be transmitted vertically. Sick sheep and resistant sheep are the main sources of infection of the disease (He Ying et al., 2009; Wan Yiyuan et al., 2000). The average incubation period of MPGS is 18-20d, the fastest is 3-6d, and the longest is 30-40d. Its clinical features are mainly high fever, cough, progressive weight loss, serous and fibrinous inflammation of lun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/04C12R1/35
CPCC12Q1/689Y02A50/30
Inventor 林裕胜胡奇林游伟江锦秀张靖鹏
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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