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Detection kit for human parainfluenzavirus3 (HPIV3) IgM antibody

A technology of human parainfluenza virus and detection kit, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of easy existence of errors, low degree of automation, and need for visual observation, and achieves the effect of eliminating false positives and automatic detection.

Inactive Publication Date: 2019-11-26
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The indirect immunofluorescence method is to fix the antigen on the glass slide, use fluorescein FITC as a marker, and observe the results through a fluorescent microscope. The interpretation of the results basically requires visual observation, which is prone to errors
Enzyme-linked immunoassay is to immobilize antigens or antibodies on polystyrene microwell plates, use horseradish peroxidase or alkaline phosphatase as markers, and judge the results by color absorbance value. The reaction time of the experiment is long and it is automated. low level
Colloidal gold chromatography immobilizes antigens or antibodies on nitrocellulose membranes, uses colloidal gold as a marker, and develops color on the detection line through liquid capillary action. The interpretation of the results basically requires visual observation, which is fast but has low accuracy.

Method used

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  • Detection kit for human parainfluenzavirus3 (HPIV3) IgM antibody
  • Detection kit for human parainfluenzavirus3 (HPIV3) IgM antibody
  • Detection kit for human parainfluenzavirus3 (HPIV3) IgM antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Preparation of human parainfluenza virus type 3 IgM antibody detection kit

[0036] 1. Preparation of anti-human IgM antibody-coupled magnetic particles

[0037] 1.1 Magnetic particle washing

[0038] Take 200 μL of magnetic particles containing carboxyl groups on the surface and put them in a glass bottle, use a magnet to adsorb the magnetic particles to the bottom of the glass bottle, remove the supernatant; add 2 mL of 0.02M PBS (pH 8.0), and repeat the above operation 3 times.

[0039] 1.2 Activation of Magnetic Particles

[0040] Dissolve EDC and NHS in 0.1M MES (pH 5.0) buffer solution at a concentration of 10-70mg / mL, then add 1mL each to the magnetic particles; shake gently at room temperature for 30-60 minutes; The particles are adsorbed at the bottom, and the supernatant is removed; then 2 mL of 0.1M MES (pH 5.0) buffer solution is added to resuspend the magnetic particles; the above operation is repeated twice.

[0041] 1.3 Conjugated anti-human ...

Embodiment 2

[0061] The detection method of embodiment 2 kit of the present invention

[0062] The kit prepared in Example 1 is used in conjunction with the automatic chemiluminescence instrument AutoLumo A2000 or AutoLumo A2000 Plus produced by Zhengzhou Antu Bioengineering Co., Ltd. to detect human parainfluenza virus type 3 IgM antibodies. The specific steps are:

[0063] 1. Add 3 wells of positive control (for determining the Cutoff value) and 2 wells of negative control in sequence in the reaction container (hereinafter referred to as "well"), 100 μL / well; add 10 μL sample and 100 μL sample diluent to each well of the remaining wells ;

[0064] 2. Add 20 μL of magnetic particle suspension to each well; after mixing, incubate at 37°C for 15 minutes; then wash 5 times with cleaning solution;

[0065] 3. Add 50 μL of enzyme conjugate and 50 μL of antigen solution to each well; mix well and incubate at 37°C for 17 minutes; wash with cleaning solution 5 times;

[0066] 4. Finally, add 50...

Embodiment 3

[0070] The performance evaluation of embodiment 3 kit of the present invention

[0071] 1. Repeatability

[0072] Repeatability refers to the repeated measurement of the same sample, and the closeness of each measurement result to the mean value, expressed in the coefficient of variation CV.

[0073] Select 3 samples with different concentrations, test 4 repetitions every day, and test continuously for 5 days, the coefficient of variation CV<8%, the results are shown in Table 1:

[0074] Table 1

[0075]

[0076] From the results shown in Table 1, it can be seen that the coefficient of variation of sample 1 is 2.91%, the coefficient of variation of sample 2 is 5.07%, and the coefficient of variation of sample 3 is 6.54%, all of which are less than the 10% required by the industry, which proves that its repeatability is good .

[0077] 2. Specificity

[0078] Eight pathogens: Mycoplasma pneumoniae, Chlamydia pneumoniae, Legionella pneumophila, adenovirus, enterovirus typ...

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Abstract

The invention relates to the technical field of in-vitro diagnosis, and especially relates to a detection kit for a human parainfluenzavirus3 (HPIV3) IgM antibody. The detection kit comprises an anti-human IgM antibody-magnetic particle conjugate, an HPIV3 antigen, a horseradish peroxidase labeled antibody of the HPIV3 antigen, a sample diluent and a chemiluminiscence substrate, wherein the enzyme-labeled antibody is an antibody without an Fc end. The detection kit adopts a capture method and is high in sensitivity and specificity. In addition, the capture method uses the natural HPIV3 as a bridge and uses the monoclonal antibody of the HPIV3 as a tracer, so that the problem that the specificity and the sensitivity are influenced due to insufficient purity of the antigen or incomplete sites when the antigen is used as the tracer is solved. The Fc end of the monoclonal antibody of the HPIV3 is cut off, so that false positive caused by a heterophile antibody existing in a sample can be eliminated.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a detection kit for human parainfluenza virus type 3 IgM antibody. Background technique [0002] Human parainfluenzavirus (HPIV) is a member of the Paramyxoviridae family and is a single-stranded negative-sense RNA virus. Four serotypes of HPIV have been identified, namely types 1, 2, 3 and 4. HPIV1 and HPIV3 belong to the Respirovirus genus, and HPIV2 and HPIV4 belong to the Mumps virus genus. Human parainfluenza virus type 3 occurs every year, and it is popular in spring and summer, but it can be popular all year round in temperate regions. Transmission is by aerosols of large droplets and contact with contaminated surfaces, with an incubation period of 2-6 days after infection. Human parainfluenza virus type 3 is an important viral pathogen that causes respiratory tract infection in infants and young children. The main clinical symptoms are laryngitis, bronchiol...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N33/543G01N33/569
CPCG01N33/535G01N33/54326G01N33/56983
Inventor 陶占领孙培育郑亚平周伟伟刘功成渠海付光宇
Owner AUTOBIO DIAGNOSTICS CO LTD
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