Chemiluminescence immunoassay kit for Seneca valley virus structural protein VP1 antibody detection

A chemiluminescence immunoassay and structural protein technology, which is applied in the field of chemiluminescence immunoassay kits, can solve the problems of short detection time and achieve the effects of short detection time, high protein yield and simple operation

Inactive Publication Date: 2018-11-23
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

[0006] The object of the present invention is to provide a kind of detection time is short, the chemiluminescence immunoassay kit that is used for Seneca Valley virus structural protein VP1 antibody detection with good specificity and high sensitivity, a...

Method used

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  • Chemiluminescence immunoassay kit for Seneca valley virus structural protein VP1 antibody detection
  • Chemiluminescence immunoassay kit for Seneca valley virus structural protein VP1 antibody detection
  • Chemiluminescence immunoassay kit for Seneca valley virus structural protein VP1 antibody detection

Examples

Experimental program
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Effect test

Embodiment 1

[0049] Example 1 Preparation and assembly of chemiluminescent immunoassay kit for detection of Seneca Valley virus structural protein VP1 antibody

[0050] (1) Preparation of the kit:

[0051] 1. Expression of Seneca Valley virus structural protein VP1 antigen:

[0052] The 792bp gene encoding the VP1 protein of Seneca Valley virus (shown in SEQ ID NO.1) was cloned into the pET-30a(+) prokaryotic expression vector. After digestion and sequencing, the positive plasmid was transformed into the BL21DE3plysS strain, Kana Screen the single clone on the resistant LB plate, shake the LB medium overnight at 37°C; transfer the overnight bacterial solution to fresh LB medium at a volume ratio (v / v) of 1:100, shake at 200rpm at 37°C for 3 hours to reach the OD600 value When it reaches about 0.6, add IPTG with a final concentration of 1mmol / L to continue shaking the bacteria for 3h, and collect the bacterial liquid by centrifugation at 6000rpm. Reduce the culture temperature to 30°C; ad...

Embodiment 2

[0089] Example 2 Sensitivity and specificity experiments of the chemiluminescent immunoassay kit for detection of Seneca Valley virus structural protein VP1 antibody

[0090] 1. Dilution of known background serum:

[0091] (1) A total of 30 serum samples were obtained from clinically healthy pigs that had not been infected with Seneca Valley virus. These sera were used to evaluate the specificity of the chemiluminescence detection method for antibodies to Seneca Valley virus structural proteins.

[0092] (2) 30 porcine vesicular disease positive sera, 30 porcine circovirus positive sera and 30 porcine circovirus-2 positive sera each. These sera were used to evaluate the specificity of the chemiluminescent detection method for Seneca Valley virus structural protein antibodies.

[0093] (3) A total of 80 serum samples were obtained from pigs experimentally challenged with Seneca Valley virus. The pigs had not received any immunizations prior to the challenge, and these serum sa...

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Abstract

The invention discloses a chemiluminescence immunoassay kit for Seneca valley virus structural protein VP1 antibody detection. The kit comprises a Seneca valley virus structural protein VP1 antigen coated chemiluminescence immunoassay plate, positive control serum, negative control serum, an enzyme-labeled antibody, chemiluminescence substrate liquid, a luminescence enhancing agent and concentrated washing liquid, wherein the amino acid sequence of the Seneca valley virus structural protein VP1 is shown as SEQ ID NO.2. The kit provided by the invention uses structural protein VP1 antigen expressed by a prokaryotic expression system; the protein yield is high; the antigen purity reaches 90 percent or higher. An enzymatic chemiluminescence reaction system is used for judging the result; thedetection sensitivity is improved. The kit provided by the invention has the advantages that the operation is simple; the detection time is short; the sensitivity and the specificity are high, and thelike. The kit can be used for Seneca valley virus detection and epidemiological investigation of a great number of samples, and has good market prospects.

Description

technical field [0001] The invention relates to a chemiluminescent immunoassay kit for detecting the antibody to the structural protein VP1 of Seneca Valley virus, and also relates to a method for using the kit. The invention belongs to the technical field of virus detection. Background technique [0002] Porcine Seneca Valley disease is a primary herpetic disease of porcine caused by Seneca Valley virus (SVV). The disease mainly causes blisters and ulcers on the mucous membranes of the snout and nose and hooves of pigs. In 2008 and 2012, SVV was diagnosed in diseased pig herds in parts of the United States and Canada. In September 2015, lameness, vesicles, and death of newborn piglets occurred in a certain pig farm in the United States. PCR detection ruled out foot-and-mouth disease virus ( FMDV), porcine epidemic diarrhea virus (PEDV), porcine rotavirus, porcine transmissible gastroenteritis virus (TGEV) and porcine respiratory and reproductive syndrome virus (PRRSV), se...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/68G01N33/535G01N21/76
CPCG01N21/763G01N33/535G01N33/56994G01N33/6854G01N2333/03G01N2333/47
Inventor 吕建亮潘丽张中旺张永光卢田鑫李姚尧
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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