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Luchuan pig mycoplasma pneumoniae P36 gene in-situ hybridization detection method, and probes and kit thereof

A technology of Mycoplasma pneumoniae and detection kit, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as the research on the location of the inability to infect the process, and achieve high sensitivity and accurate detection and prediction. , a strong specific effect

Inactive Publication Date: 2015-09-09
GUANGXI VETERINARY RES INST +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection of Mhp in Luchuan pigs in Guangxi mainly uses the PCR detection method, but this method is only limited to the detection of pathogens, and cannot be used to locate the infection process of Mhp

Method used

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  • Luchuan pig mycoplasma pneumoniae P36 gene in-situ hybridization detection method, and probes and kit thereof
  • Luchuan pig mycoplasma pneumoniae P36 gene in-situ hybridization detection method, and probes and kit thereof
  • Luchuan pig mycoplasma pneumoniae P36 gene in-situ hybridization detection method, and probes and kit thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0028] 1. Specific oligonucleotide probes and labels

[0029] According to the three specific DNA sequences in the whole gene fragment of Mycoplasma hyopneumoniae P36 (X67286, 1394bp) in Luchuan, the corresponding base regions are 417-452, 516-551, and 687-722, respectively, and three oligonucleotides were designed and synthesized Probes were synthesized and labeled with digoxigenin DIG at the 3' end of each probe.

[0030] 2. Establishment of in situ hybridization detection method for Mycoplasma hyopneumoniae P36 gene in Luchuan

[0031] 1. Test material

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Abstract

The invention discloses a Luchuan pig mycoplasma pneumoniae P36 gene in-situ hybridization detection method, and probes and a kit thereof. Three oligonucleotide probes are designed and synthesized according to three specific DNA (deoxyribonucleic acid) sequences in the Luchuan pig mycoplasma pneumoniae P36 whole-genome segment. The invention also discloses a related detection method and kit. The specific probes have high specificity and high sensitivity. The detection method and kit established on such basis can be conveniently and quickly used for digoxin labeled in-situ hybridization detection of Luchuan pig mycoplasma pneumoniae P36 gene. The method can quickly and accurately identify Luchuan pig mycoplasma pneumoniae when being used for detection and monitoring research of Gangxi Luchuan pig mycoplasma pneumoniae, thereby determining the distribution conditions of the mycoplasma pneumoniae DNA in the Luchuan pig lung tissues. The Luchuan pig mycoplasma pneumoniae pathogen nucleic acid distribution conditions are combined with the related pathology tissue changes to achieve the goal of accurate detection and prediction.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Luchuan mycoplasma hyopneumoniae P36 gene in situ hybridization detection method, a probe and a kit thereof. Background technique [0002] Mycoplasma pneumoniae of swine or mycoplasma pneumoniae of swine (Mycoplasma pneumoniae of Swine, MPS) is a kind of chronic respiratory tract contagious disease, the causative agent of this disease is mycoplasma hyopneumoniae (Mycoplasma hyopneumoniae, Mhp) Bacillus, Actinobacillus pleuropneumoniae, porcine reproductive and respiratory syndrome virus, swine influenza virus and other synergistic porcine respiratory syndrome, a serious threat to the modern pig industry, is the main cause of relatively high mortality. Natural infection is only seen in pigs, and all kinds of pigs can be infected regardless of age, sex, and breed. Luchuan pig has the characteristics of thin skin, tender meat, crisp but not greasy, etc. It is one of the e...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68C12Q1/04
Inventor 韦英益何颖陈忠伟赵武黎珂钰秦毅斌卢冰霞梁家幸李斌苏乾莲杨思仪周英宁蒋冬福卢定专
Owner GUANGXI VETERINARY RES INST
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