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Mycoplasma capricolum subsp. capripneumoniae specific protein and indirect ELISA kit

A mycoplasma, specific technology, applied in biological testing, biochemical equipment and methods, material testing products, etc., can solve problems such as no reports of candidate antigens

Active Publication Date: 2020-06-30
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These can be used as Mccp-specific antigen candidate targets, but there is no report indicating whether these candidate antigens can be further applied

Method used

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  • Mycoplasma capricolum subsp. capripneumoniae specific protein and indirect ELISA kit
  • Mycoplasma capricolum subsp. capripneumoniae specific protein and indirect ELISA kit
  • Mycoplasma capricolum subsp. capripneumoniae specific protein and indirect ELISA kit

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Embodiment Construction

[0014] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0015] 1. Experimental materials

[0016] Main reagents: plasmid pET-30a, competent BL 21 (DE3), LB, LA, IPTG, QIGEN resin, skimmed milk powder, secondary antibody (rabbit anti-goat, sigma), Mccp antibody detection cELISA kit (IDEXX company), BCA kit, urea.

[0017] Main instruments: shaker, incubator, ultrasonic breaker, centrifuge, protein electrophoresis instrument, microplate reader. Serum: collected at different time points in the vaccine / infection experiment.

[0018] 2. Sequence acquisition

[0019] BLAST the genome sequence of Mccp 1601 (NZ_CP017125.1) with the genome sequences of other...

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Abstract

The invention discloses mycoplasma capricolum subsp. capripneumoniae specific protein and an indirect ELISA kit. The protein 87.1 has a nucleotide sequence as shown in SEQ ID NO.1. The indirect ELISAantibody detection kit prepared from the mycoplasma capricolum subsp. capripneumoniae specific protein 87.1 comprises an antigen protein 87.1 coated ELISA plate, sample diluent, positive serum, negative serum, washing liquid, confining liquid, a rabbit anti-goat secondary antibody, substrate color developing liquid and a stop solution. The coating amount of the antigen protein 87.1 is 625 ng / hole,and the dilution degrees of the positive serum and the negative serum are both 1 / 100. The indirect ELISA kit prepared from the Mccp specific protein 87.1 provided by the invention has good specificity, reacts with Mccp positive serum and does not react with Mmc and Mcc positive serums.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a mycoplasma capricum subspecies-specific protein and an indirect ELISA kit. Background technique [0002] Goat contagious caprine pleuropneumonia (CCPP) is a severe respiratory disease of goats caused by Mycoplasma capricolumsubsp. one of infectious diseases. Mccp belongs to the Mycoplasma mycoides cluster, and members of this cluster have high serological and genomic similarities. In addition to Mccp, it also includes: Mycoides subsp. mycoides (Mmm), Mycoides goat Subspecies (M.mycoidessubsp.capri, Mmc), Mycoplasma capricosum subspecies (M.capricolumsubsp.Capricolum, Mcc), Mycoplasma leechii (M.Leachii, Ml), wherein Mmm and Ml are Mycoplasma bovis, Mmc, Mcc and Mccp are mycoplasma of sheep origin, both of which can cause sheep pneumonia. The closest relatives to Mccp are Mcc and Ml, which are prone to cross-reaction with Mccp in serological detection. Antigens are t...

Claims

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Application Information

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IPC IPC(8): C12N15/31G01N33/68G01N33/569G01N33/543
CPCC07K14/30G01N33/6854G01N33/56933G01N33/54393Y02A50/30
Inventor 储岳峰吴娅琴颜新敏郝华芳陈胜利马丽娜刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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