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ELISA kit for detecting Brucella antibody

The technology of a brucella and a kit is applied in the field of ELISA kits for detecting brucella antibodies, which can solve the problems of a large number of reagents required, a long detection cycle, complicated procedures, etc., and achieve low detection costs and are suitable for promotion , the effect of high sensitivity

Inactive Publication Date: 2019-03-22
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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Problems solved by technology

The traditional detection method of Brucella is far from meeting the requirements of modern detection due to its shortcomings such as long detection cycle, complicated procedures, and various reagents required.
There are some problems in the practical application of pathogenic nucleic acid detection technology represented by polymerase chain reaction technology. For example, ordinary polymerase chain reaction technology requires special instruments, and has the characteristics of easy cross-contamination and cumbersome operation process.

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  • ELISA kit for detecting Brucella antibody
  • ELISA kit for detecting Brucella antibody
  • ELISA kit for detecting Brucella antibody

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Embodiment Construction

[0010] The present invention will be further described in detail below in conjunction with specific embodiments:

[0011] The ELISA kit for detecting Brucella antibodies of the present invention includes an Omp31 recombinant protein coated plate. The Omp31 recombinant protein uses a part of the Omp31 gene sequence and is codon-optimized, as shown in SEQ ID NO. 1 in the sequence table. The nucleotide sequence shows the complete gene synthesis. After digestion, the Omp 31 gene was ligated into the pET-28a(+) plasmid, and after identification, it was transformed into E. coli BL21 competent cells, induced expression, and obtained by affinity chromatography. of.

[0012] The coating method of the protein-coated plate is as follows: Omp31 recombinant protein is diluted with 0.5M carbonate buffer, 0.1 mL is added to each well of a 96-well microtiter plate, and coated at 2-8°C for 15 hours; 250 μL is added to each well The sealing solution is sealed at room temperature for 2 hours, and th...

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Abstract

The invention discloses an ELISA kit for detecting a Brucella antibody. An Omp31 gene is used for constructing recombinant engineering bacteria, an Omp31 expression recombinant protein is used as a coated antigen for preparing a protein coated plate, and the ELISA antibody detecting kit is established. The kit is good in specificity, high in sensibility, low in detection cost, and suitable for popularization. An important method is provided for a disease in aspects of antibody level detection, epidemiological investigation, similar disease identification and effective control measure adoptionand the like. The kit comprises the following main components: the Omp31 protein coated plate, positive control serum, negative control serum, an enzyme labeling rabbit anti-bovine polyclonal antibody, substrate liquid, a stop solution, a diluent, washing liquid, and a serologecal plate.

Description

Technical field [0001] The invention relates to an ELISA kit, in particular to an ELISA kit for detecting Brucella antibodies. Background technique [0002] Brucella is a gram-negative, intracellular parasitic bacterium. It is a small rod-shaped bacterium that can survive in many kinds of livestock. Cattle, sheep, pigs, dogs, camels, deer and other animals may be infected. Contact with infected animals or infected food and laboratory contact can spread to humans. The main causes of human infection are Brucella malta ("Malta fever"), Brucella abortus, Brucella bovine, Ovis, Brucella suis and Brucella canis, among which Brucella sheep Bacteriosis is the most common. [0003] In 1886, British military doctor Bruce isolated "Brucella" from the spleens of soldiers who died of "Maltese fever" on the island of Malta, and for the first time identified the pathogen of the disease. After a person gets sick, the first symptom that appears is fever. The body temperature can reach 38-40°C fo...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/535G01N33/531
CPCG01N33/531G01N33/535G01N33/6893G01N2800/26
Inventor 肖妍霍蕾赵祥平董志珍赵丹王建华张俊哲马立才刘河冰杨柳李蓉蓉
Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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