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Real-time fluorescent quantitative PCR detection primer used for mycoplasma capricolum subsp.capripneumonia

A technology for real-time fluorescence quantification and detection of primers, applied in the field of molecular biology, can solve problems such as unseen detection, and achieve the effects of high sensitivity, good repeatability and good specificity

Inactive Publication Date: 2017-07-21
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After reviewing domestic and foreign literature, it was found that no SYBR Green Ⅰ real-time fluorescent quantitative PCR method for detecting Mccp has been found so far.

Method used

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  • Real-time fluorescent quantitative PCR detection primer used for mycoplasma capricolum subsp.capripneumonia
  • Real-time fluorescent quantitative PCR detection primer used for mycoplasma capricolum subsp.capripneumonia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Establishment of Mccp SYBR Green Ⅰ fluorescence quantitative PCR method

[0020] 1. Materials:

[0021] Mccp F38 strain was donated by the Lanzhou Research Institute of the Chinese Academy of Sciences, and Escherichia coli, Pasteurella, Staphylococcus aureus, Haemophilus parasuis, etc. were donated by the Poultry Disease Laboratory and the Pig Disease Laboratory of the Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences; Mycoplasma pneumoniae and aphthus virus were isolated and preserved in our department.

[0022] Two, steps

[0023] 1. Instruments and reagents:

[0024] Mastercycler ep realplex fluorescence quantitative PCR instrument was purchased from eppendorf; gel recovery kit was purchased from Corning Life Science (Wujiang) Co., Ltd.; SYBR Premix Ex Taq II (2×), PMD19-T vector, and DL2000Marker were purchased from Treasure Bioengineering (Dalian) Co., Ltd.; plasmid mini-extraction kit was purchased from Om...

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Abstract

The invention provides a real-time fluorescent quantitative PCR detection primer used for mycoplasma capricolum subsp.capripneumonia. The primer sequences are as follows: the sequence of a forward primer is 5'-GAACTGAAGAAGGTATGGCTGAAGA-3', and the sequence of a reverse primer is 5'-CCTGTTCCAGCACCAACTAAAAC-3'. According to the arcD gene sequence of AY529462.1 logged in the GenBank, the specific primer is designed, a SYBR Green I real-time fluorescence quantitative PCR method of the mycoplasma capricolum subsp.capripneumonia is firstly built at home and abroad, and the method is good in specificity, high in sensitivity and good in repeatability.

Description

technical field [0001] The invention relates to a real-time fluorescence quantitative PCR detection primer for mycoplasma capricosum subsp. goat pneumonia, belonging to the field of molecular biology. Background technique [0002] Mycoplasma capricosum subsp. goat pneumonia (Mycoplasma capricolum subsp. capripenumoniae, Mccp) is a member of the mycoplasma mycoplasma cluster and is the cause of goat infectious pleuropneumonia ( contagious Capripleuropneumonia , CCPP) pathogen. Infected flocks have no age and gender differences, and the morbidity and mortality are high (Chu Yuefeng, 2009; Zhao Ping, 2010), and the incidence rate of the disease is 100% (Nichlas et al., 2012; Chuet al., 2011), the mortality rate was as high as 60-70% (Zheng Yingying, 2014). The incubation period of the disease is 2-28 days, with an average of 10 days, and its clinical features are mainly fibrinous pleuropneumonia. CCPP can be traced back to Algeria in 1873. In 1976, Macowan and Minette su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/689C12Q1/686C12Q2561/113C12Q2563/107C12Q2545/114
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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