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A kind of rpa test kit and application thereof for rapid detection of mycoplasma in chicken synovial bursa

A technology of Mycoplasma synovialis and kit, which is applied in 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 before, reduce the possibility of product contamination, and the method is simple , the effect of high sensitivity

Active Publication Date: 2022-02-18
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But as far as we know, there is no application of RPA technology in MS at home and abroad

Method used

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  • A kind of rpa test kit and application thereof for rapid detection of mycoplasma in chicken synovial bursa
  • A kind of rpa test kit and application thereof for rapid detection of mycoplasma in chicken synovial bursa
  • A kind of rpa test kit and application thereof for rapid detection of mycoplasma in chicken synovial bursa

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Experimental program
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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 an RPA kit and a detection method for rapidly detecting mycoplasma of the chicken synovial bursa. The kit includes: a reaction tube containing RPA freeze-dried particles, reaction buffer, magnesium acetate, a pair of primers and a probe. Experiments have proved that the sensitivity and specificity of the kit of the present invention are equivalent to those of the qPCR method, the entire reaction time is completed within 20 minutes, and the detection result only needs to be judged by the change of the fluorescence curve. Therefore, the kit of the present invention has the advantages of quickness, high efficiency, sensitivity, etc., not only provides a technical means for effectively detecting and identifying Mycoplasma synovial bursa, but also is easy and fast to operate, and is suitable for clinical veterinary on-site diagnosis and diagnosis of chicken synovial bursa Scientific research on mycoplasma.

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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/6844C12N15/11C12R1/35
CPCC12Q1/689C12Q1/6844
Inventor 宫晓炜陈启伟郑福英刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI