Kit for quantitative detection on O type foot-and-mouth disease virus antibody through fluorescence immunoassay magnetic particles

A quantitative detection and fluorescence immunoassay technology, which is applied in the field of animal disease detection, can solve the problems of time-consuming operation, long detection time, cumbersome and time-consuming operation, etc., and achieve the effect of shortening the reaction time, simple and fast operation, and improving sensitivity

Inactive Publication Date: 2018-06-01
GUANGZHOU BIOKEY HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In serological diagnostic techniques, the standard method for detecting FMDV antibodies recommended by OIE is neutralization test, but the neutralization test needs to use live virus, which needs to be completed in a special laboratory, and the operation is time-consuming
Enzyme-linked immunosorbent assay has the advantages of sensitivity and high specificity, but the operation is cumbersome and time-consuming
Chemiluminescence method has the advantages of high sensitivity and good specificity, but it needs special, large and expensive equipment, and the detection time is long

Method used

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  • Kit for quantitative detection on O type foot-and-mouth disease virus antibody through fluorescence immunoassay magnetic particles
  • Kit for quantitative detection on O type foot-and-mouth disease virus antibody through fluorescence immunoassay magnetic particles

Examples

Experimental program
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Effect test

Embodiment 1

[0032] In this embodiment, the kit for quantitative detection of type O foot-and-mouth disease antibody by fluorescent immunomagnetic particles consists of a reagent strip and type O foot-and-mouth disease negative and positive serum.

[0033] Among them, the reagent strip consists of test well 1 (VP1 protein coated carboxyl magnetic beads), well 3 (biotinylated goat anti-pig secondary antibody), well 4 (streptavidin-labeled europium), washing liquid well 6 and 7. Strengthen the composition of liquid hole 12, such as figure 1 with figure 2 Shown.

[0034] The preparation process of the fluorescent immunomagnetic particle quantitative detection kit for O-type foot-and-mouth disease antibody is as follows:

[0035] (1) Preparation method of immunomagnetic beads:

[0036] Washing magnetic beads

[0037] Pipette 1mg of carboxyl magnetic beads with a diameter of 1μm into a 1.5ml centrifuge tube, place the centrifuge tube in a magnetic stand, use the magnetic stand to separate the magneti...

Embodiment 2

[0072] Example 2 Preparation and detection of reagent strips

[0073] The reagent strips in this example are semi-finished products assembled through the following processes: Dispense 100 μL of immunomagnetic beads, 200 μL of biotinylated goat anti-pig antibody, SA-Eu into the first, third, fourth, sixth, seventh, and twelfth stage. 3+ , 900μL lotion, 900μL lotion, 300μL enhancement solution, and then sealed with a film sealing machine. The sealing film is coated with product information identification that can be scanned and identified by the automatic fluorescence detection analyzer, including corporate standard curves, batches , Production date, expiration date. The finished reagent strip, the pre-packed O-type foot-and-mouth disease negative serum, positive serum and other accessories are assembled into a kit.

[0074] Sample testing:

[0075] ①Add sample:

[0076] Put the sample to be tested or the O-type foot-and-mouth disease antibody standard into the loading system of the au...

Embodiment 3

[0090] Example 3 Sensitivity detection

[0091] The steps of this method are as follows:

[0092] (1) Dilute the porcine type O FMDV positive standard serum with a calibrated antibody titer of 1:512 from 1:32 to 1:32, 1:64, 1:128, 1 : 256, 1:512, 1:1024 six dilutions, put the diluted sample into the sample compartment.

[0093] (2) Add 100μl of biotinylated goat anti-pig antibody to the test well of the reagent strip, and incubate at 37°C for 10min; then discard the supernatant, wash with washing solution 5 times, adding 200μl washing solution each time;

[0094] (3) Add 100μl SA-Eu diluted 1:80000 to the test hole of the reagent strip 3+ , Incubate at 37°C for 10 minutes, then discard the supernatant and wash with lotion;

[0095] (4) Add 100μl of enhancement solution to the test hole of the reagent strip and incubate for 5 minutes;

[0096] (5) The detector automatically detects the fluorescence value.

[0097] If the fluorescence value of the serum to be tested is ≥ 37695, the serum t...

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Abstract

The invention discloses a kit for quantitative detection on an O type foot-and-mouth disease virus antibody through fluorescence immunoassay magnetic particles. The kit consists of O type foot-and-mouth disease virus antibody negative serum, O type foot-and-mouth disease virus antibody positive serum, VP1 coating magnetic beads, a biotinylation goat-anti-pig antibody, a streptavidin marking fluorescent substance, a cleaning solution and an enhancing solution. The magnetic beads used in the kit have relatively large binding areas, so that the detection range is greatly increased, the reaction time is shortened, and the sensitivity is improved. The kit has a relatively wide stimulation spectrum and a relatively narrow emitting spectrum, the cost can be reduced, and the sensitivity can be improved; compared with a conventional fluorescent substance, the kit is relatively wide in detection range and relatively good in specificity. Due to adoption of a streptavidin-biotin signal amplification system, the detection sensitivity is further improved, and the kit is relatively high in sensitivity when being compared with ELISA (Enzyme-Linked Immuno Sorbent Assay) and chemiluminiscence. Together with a full-automatic detector, on-site automatic operation can be achieved, one or more samples can be simultaneously detected, and the kit is simple, convenient and rapid to operate and low in price.

Description

Technical field [0001] The present invention relates to the technical field of animal disease detection. Specifically, the present invention relates to a kit for the quantitative detection of type O foot-and-mouth disease antibodies by fluorescent immunomagnetic particles. Background technique [0002] Foot-and-mouth disease virus (Foot-and-mouth disease virus, FMDV) can cause acute, febrile, and highly contact infectious diseases in cloven-hoofed animals, which is commonly referred to as Foot and Mouth Disease (FMD). The disease caused by the virus has the characteristics of rapid spread and high infection rate. It mainly spreads through the respiratory and digestive tracts. Only a few infectious virus particles can cause the disease in susceptible animals, and even cause a worldwide pandemic, bringing to the animal husbandry industry. Because of the serious economic losses, FMDV is the key target of quarantine and epidemic prevention in all countries in the world. FMDV is a s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/531G01N33/533
CPCG01N33/6857G01N33/531G01N33/533G01N33/54326
Inventor 李根平
Owner GUANGZHOU BIOKEY HEALTH TECH CO LTD
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