RPA kit for rapidly detecting mycoplasma synoviae of chicken and its use

A technology of Mycoplasma synovialis and kit, which is applied in the directions of DNA/RNA fragments, recombinant DNA technology, microorganism-based methods, etc., can solve problems such as the application of RPA technology that has not been seen, and achieves reduced possibility, strong specificity, and increased specific effect

Active Publication Date: 2019-10-08
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • RPA kit for rapidly detecting mycoplasma synoviae of chicken and its use
  • RPA kit for rapidly detecting mycoplasma synoviae of chicken and its use
  • RPA kit for rapidly detecting mycoplasma synoviae of chicken and its use

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Embodiment 1

[0033] Embodiment 1 The establishment of a kind of RPA kit and detection method of rapidly diagnosing mycoplasma chicken synovial bursa

[0034] 1. Design and preparation of primers and probe sequences

[0035] The present invention designs primers and probes according to the hypothetical protein VY93_00965 gene of Mycoplasma gallinae bursa; through the comparison and analysis of Blast software in NCBI, it is found that this gene has a high degree of conservation with different strains of Mycoplasma gallisovus uploaded in GenBank The laboratory found that the gene exists in different clinical isolates through proteome sequencing, and the results of amplification and sequencing confirmed that the homology of the gene was as high as 98.5%. Therefore, the present invention selects a section (701bp-920bp) in the full length of the gene as the target fragment to design primers and probes, in order to detect as many Mycoplasma gallisovus as possible. All primers and probes were syn...

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Abstract

The invention discloses a RPA kit for rapidly detecting mycoplasma synoviae of chicken and its use. The kit comprises: a reaction tube containing RPA lyophilized particles, a reaction buffer solution,magnesium acetate, a pair of primers and a probe. The experiments prove that the sensitivity and specificity of the kit of the present invention are comparable to those of a qPCR method, and the entire reaction time is completed within 20 minutes, and the detection result only needs to be determined by the change of a fluorescence curve. Therefore, the kit of the invention has the advantages of quickness, high efficiency, sensitivity, and the like, provides a technical means for effectively detecting and identifying the mycoplasma synoviae of chicken, is easy and fast to operate, and is suitable for clinical veterinary on-site diagnosis and scientific research on mycoplasma synoviae of chicken.

Description

technical field [0001] The invention belongs to the specific field of preventive veterinary examination, relates to a kit for detecting Mycoplasma bursa and its application, in particular to an RPA detection kit for rapidly detecting Mycoplasma bursa and its application. Background technique [0002] Mycoplasma synoviae (MS) belongs to Mycoplasma order, Mycoplasma family, and Mycoplasma genus. It is an important pathogen that infects chickens and has brought serious economic losses to the poultry industry in my country. Chicken flocks infected with MS can cause upper respiratory tract disease, air sacculitis, joint synovial bursitis and tenosynovitis, which are mainly manifested clinically as joint swelling, lameness, growth retardation of chicks and decreased egg production of laying hens; It can spread vertically and can spread among chickens; since 2008, MS infection has been detected in farms in many provinces and regions of my country; when MS is combined with other path...

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6844C12N15/11C12R1/35
CPCC12Q1/689C12Q1/6844C12Q2521/507C12Q2522/101
Inventor 宫晓炜陈启伟郑福英刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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