Rapid detecting mycoplasma ovipneumoniae method

A technology of mycoplasma pneumoniae and sheep, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection. It can solve the problems of PCR method sensitivity, dependence on precision instruments, and inability to directly detect pathogens, etc., and achieve high specificity. performance and sensitivity, and the effectiveness of accurate detection methods

Inactive Publication Date: 2017-02-22
SHIHEZI UNIVERSITY
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Problems solved by technology

Indirect ELISA has high specificity, sensitivity, stability and reproducibility, but it can only detect MO antibody and cannot directly detect pathogens
[0009] The PCR method is sensitive and fast, but it relies on expensive and complex precision instruments, maki

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  • Rapid detecting mycoplasma ovipneumoniae method
  • Rapid detecting mycoplasma ovipneumoniae method
  • Rapid detecting mycoplasma ovipneumoniae method

Examples

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[0033] In this example, sheep from pastures in the Xinjiang Uygur Autonomous Region were used as specimens to aseptically cultivate and collect sheep Eg head sections, and other raw materials were obtained by purchasing kits.

[0034] 1. Design of LAMP primer

[0035] According to the MO 16S rDNA sequence, DNAMAN software was used for sequence alignment, and the highly conserved MO sequence was selected as the target sequence for amplification.

[0036] According to the primer design principle and reaction principle of LAMP technology, multiple primer pairs are designed and generated in the Primer ExplorerV4 software system, and suitable primer pairs are screened for primer synthesis. The primer pair designed in this research includes a pair of inner primers (FIP-BIP) and a pair of outer primers (F3-B3). The length of the amplified target gene fragment is 188bp. The primer names and sequences are shown in Table 1.

[0037] Table 1 Name and sequence of LAMP specific primers

[0038]

[...

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Abstract

The invention relates to a rapid detecting mycoplasma ovipneumoniae method, according to the gene sequence of the detected mycoplasma ovipneumoniae designing a LAMP primer; establishing a reaction system for LAMP visualization method, and conducting optimizations for LAMP reaction conditions; gathering and embrocating the sheep nasal secretions, preparing DNA template solution, conducting detections using the optimized reaction system and reaction condition. The method targets the shortcomings of requiring professional instruments and labs and the like in detecting mycoplasma ovipneumoniae in the existing methods, which limits the promotion and application of these diagnosis methods in clinics, and establishes a rapid detecting LAMP visualization method to detect MO, the method has very high specificity and sensitivity, and provides a rapid, simple, and accurate detection method for clinical diagnoses of sheep MO infection.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a method for rapidly detecting ovine mycoplasma pneumoniae in a visualized manner. Background technique [0002] Mycoplasma ovis pneumonia, also known as ovine infectious pleuropneumonia, is caused by Mycoplasma ovis pneumoniae (Mycoplasma ovipneumoniae, MO) causing interstitial, atypical pneumonia, and suppurative pneumonia in sheep and goats. [0003] Since the first report in sheep by Mackay et al. in 1963, Mycoplasma ovis pneumonia has been widespread worldwide. MO not only infects sheep and goats, but also other ruminants such as bighorn sheep. In recent years, there have been reports of the occurrence and prevalence of Mycoplasma ovis pneumonia in Xinjiang, Gansu, Inner Mongolia, Qinghai, Shaanxi, Ningxia, Liaoning, Jiangsu, Sichuan, Yunnan and other provinces with developed sheep farming industry. With the healthy development of the sheep industry, the harm i...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/35
CPCC12Q1/6844C12Q2531/119
Inventor 乔军孟庆玲田路路卢海亭乔梦凡贡莎莎陈诚李重阳孟丹于伟伟陈创夫
Owner SHIHEZI UNIVERSITY
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