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Primer and probe for detecting Mycoplasma ovipneumoniae

A technology for Mycoplasma pneumoniae and sheep, which is applied in the field of molecular biology and can solve problems such as Mycoplasma ovine pneumonia that have not yet been seen

Pending Publication Date: 2017-09-08
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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Problems solved by technology

Through reviewing the literature, it was found that although there is a real-time fluorescent quantitative PCR method for the detection of Mycoplasma pneumoniae in China, it is mainly based on primers designed for 16S rRNA, and there is only a dye-based fluorescent quantitative PCR method, and Mycoplasma pneumoniae has not been seen yet. Taq Man fluorescent quantitative PCR detection method

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  • Primer and probe for detecting Mycoplasma ovipneumoniae
  • Primer and probe for detecting Mycoplasma ovipneumoniae

Examples

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Embodiment 1

[0021] Example 1 Establishment of the Fluorescent Quantitative PCR Method for Mycoplasma Ovis Pneumoniae

[0022] 1. Materials:

[0023] Mycoplasma goat pneumonia subspecies was donated by Chu Yuefeng, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences; E. FJ-CL01 strain, Mycoplasma goat subspecies FJ-GT strain, Acholesteroplasma reinhardt FJ-NP strain, Mycoplasma bovis FJ-HJ strain, Oral ulcer virus FJ-FZ strain were isolated, identified and preserved in our laboratory .

[0024] Two, steps

[0025] 1. Instruments and reagents: pMD19-T vector, SYBR Premix Ex Taq Ⅱ (2×), DL2000 Marker were purchased from Treasure Bioengineering (Dalian) Co., Ltd.; fluorescence quantitative PCR tube was purchased from Axygen (USA); gel recovery kit was purchased from Corning Life Sciences (Wujiang) Co., Ltd.; Extraction kits were purchased from Omega Bio-Tek (USA); Mastercycler ep realplex fluorescence quantitative PCR instrument, products of eppendorf (Germany...

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Abstract

The invention provides a primer and a probe for detecting Mycoplasma ovipneumoniae. A primer sequence comprises an upstream primer: 5'-CTTCGGGACTTATTGGAG-3', and a downstream primer: 5'-GATGCAAACTGATTTACTTG-3', and the probe sequence is as follows: 5'- AAGACCGATTGTCAGGCCGA-3'. The specific primer and the probe are designed according a p113 gene sequence of KR270152.1 registered in GenBank, a real-time fluorescence quantification PCR method is established aiming at the Mycoplasma ovipneumoniae p113 gene for the first time, the method has the advantages of good specificity, high sensitivity and good repeatability, and the primer and the probe can be used for detecting content of Mycoplasma ovipneumoniae in a clinic sample.

Description

technical field [0001] The invention relates to a primer and a probe for detecting sheep mycoplasma pneumoniae, belonging to the field of molecular biology. Background technique [0002] Mycoplasma ovis pneumoniae ( Mycoplasma ovipneumoniae , Mo) is the main pathogen causing Mycoplasma pneumonia of goats and sheep (MPGS) in sheep and goats. MPGS is highly contagious, mainly through respiratory tract infection, and can also be transmitted vertically. Diseased sheep and resistant sheep are the main sources of infection of the disease (He Ying et al., 2009; Wan Yiyuan et al., 2000). The clinical symptoms of MPGS are high fever, cough, panting, progressive weight loss, and serous and fibrinous inflammation of the lung and pleura. et al., 2011; Dassnayake et al., 2010; Besser et al., 2008). [0003] At present, there are reports of the disease occurring in Africa, Spain, Italy, the United States, Jordan abroad, and in Sichuan, Guizhou, Inner Mongolia, Qinghai, Guangxi, Fujian ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06C12N15/11
CPCC12Q1/6851C12Q1/689C12Q2561/101
Inventor 林裕胜胡奇林江锦秀游伟
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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