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Porcine reproductive and respiratory syndrome virus antibody detection kit and application thereof

A respiratory syndrome and antibody detection technology, applied in biological testing, measuring devices, immunoassays, etc., to achieve the effect of short time, high sensitivity, and simple operation

Pending Publication Date: 2022-06-07
禾旭(郑州)生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Each diagnostic technique has its own advantages and limitations, and the above detection methods for PRRS have deficiencies

Method used

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  • Porcine reproductive and respiratory syndrome virus antibody detection kit and application thereof
  • Porcine reproductive and respiratory syndrome virus antibody detection kit and application thereof
  • Porcine reproductive and respiratory syndrome virus antibody detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The anti-fluorescein FITC antibody-labeled magnetic microparticle solution, wherein the anti-fluorescein FITC antibody is coupled with the anti-FITC antibody by a two-step glutaraldehyde method, and contains 0.5% BSA, 1% casein, The blocking was completed with phosphate buffer at pH 7.2 (0.02 mol / L).

[0020] The preparation of the fluorescein FITC-labeled porcine reproductive and respiratory syndrome virus antigen solution includes the following steps:

[0021] S1: Mix FITC and porcine reproductive and respiratory syndrome virus at 1:1, and react at 37°C in the dark for 12 hours to form a labeling solution.

[0022] S2: dialyze the labeled solution obtained in S1 with 0.01MPBS at 4°C for 24h, add an equal volume of glycerol, and store at -20°C.

[0023] The present invention also provides an application of the porcine reproductive and respiratory syndrome virus antibody detection kit, characterized in that it includes the following steps:

[0024] S1: Put the sample ...

Embodiment 2

[0032] specific experiment

[0033] Three batches of prepared ASFV antibody magnetic particle chemiluminescence kits were used to detect ASFV antibody positive serum (20); ASFV antibody negative serum (15); swine fever virus positive serum 5; African swine fever virus positive serum 5; 5 Seneca virus positive sera; 5 foot-and-mouth disease virus O-positive sera; 5 foot-and-mouth disease virus A-positive sera; porcine vesicular virus positive sera (5); a total of 65 sera were tested, according to the measured luminescence value Calculate S / N to determine negative and positive.

[0034] Table 1 Specificity test of porcine reproductive and respiratory syndrome virus antibody detection kit

[0035]

[0036]

[0037] The results showed that the "Swine Reproductive and Respiratory Syndrome Virus Antibody Detection Kit" did not cross-react with the positive serum of swine fever, porcine reproductive and respiratory syndrome, Seneca, foot-and-mouth disease type O, foot-and-mout...

Embodiment 3

[0039] Sensitivity test

[0040] Dilute the positive standard serum with the sample diluent at 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128, 1:256, 1:512, 1:1024, It was diluted at a ratio of 1:2048 and used as a sensitive quality control serum for detection with three batches of ASFV magnetic particle chemiluminescence kits prepared.

[0041] Table 2 Sensitivity test of porcine reproductive and respiratory syndrome virus antibody detection kit

[0042]

[0043]

[0044] The test results showed that the data of the three repeated experiments were consistent, and the minimum detection amount of porcine reproductive and respiratory syndrome antibody could reach 1:256.

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Abstract

The invention relates to the technical field of immunodetection analysis, in particular to a porcine reproductive and respiratory syndrome virus antibody detection kit and application thereof. The kit takes an anti-fluorescein FITC antibody labeled magnetic particle solution, a fluorescein FITC labeled porcine reproductive and respiratory syndrome virus antigen solution, a marker, a quality control product, a luminous substrate, a buffer solution and a concentrated washing solution as components. The magnetic particles are used as carriers, so that the coating surface area and the effective coating amount of the antibody are increased, the antibody is saved, and the establishment of a relatively wide detection range is facilitated; meanwhile, an anti-fluorescein FITC antibody coupling magnetic bead is adopted to increase the antibody binding rate, so that the binding with a fluorescein-labeled biological ligand is quicker and more accurate, and non-specific binding is well reduced; according to the invention, the chemiluminescence detection technology with high sensitivity and the immunoreaction with high specificity are combined, the operation is simple, the consumed time is short, long-time color development and termination are not needed, the sensitivity is high, the specificity is good, and the porcine reproductive and respiratory syndrome virus antibody can be rapidly, simply and sensitively detected.

Description

technical field [0001] The invention relates to the technical field of immune detection and analysis, in particular to a porcine reproductive and respiratory syndrome virus antibody detection kit and application thereof. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is an infectious disease of porcine reproductive and respiratory disorders caused by porcine reproductive and respiratory syndrome virus. The sows showed miscarriage, premature birth, stillbirth, mummified fetuses and weak litters. Piglets and finishing pigs have difficulty breathing. Piglet morbidity and mortality are high and pose a major hazard to the pig industry. Specific symptoms include a significant increase in body temperature, which can reach above 41 °C; conjunctivitis and eyelid edema; respiratory symptoms such as cough and asthma; neurological symptoms such as weakness in the hindquarters, inability to stand or ataxia of some pigs; the incidence of piglets can r...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/68G01N33/543G01N33/58
CPCG01N33/56983G01N33/6854G01N33/54326G01N33/582G01N2333/08G01N2469/20
Inventor 李秀梅邵光喜刘近任雪建张和平杨晓帆杨转姬杰菲沈晓武吕园园刘明瑞殷笑丹石书霞李凯武阴思晴苏扬原鑫李卫丽王莹张静一李广玲崔亚茹王文如赵小月
Owner 禾旭(郑州)生物技术有限公司