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Antinuclear antibody quantitative detection kit and use method thereof

A technology for quantitative detection of anti-nuclear antibodies, applied in measuring devices, instruments, and material analysis through optical means, can solve problems such as cumbersome operations, reduced work efficiency, and heavy workload

Inactive Publication Date: 2015-05-20
NAT INST OF METROLOGY CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the nuclear fluorescence signal used as a quantitative indicator of indirect immunofluorescence detection has previously relied on the subjective judgment of the naked eye of the inspector, which largely caused the lack of repeatability and reliability of the quantitative results.
Moreover, the existing indirect immunofluorescence titer quantification method requires continuous gradient dilution, which is cumbersome to operate and heavy workload, resulting in reduced work efficiency and limited number of test samples

Method used

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  • Antinuclear antibody quantitative detection kit and use method thereof
  • Antinuclear antibody quantitative detection kit and use method thereof
  • Antinuclear antibody quantitative detection kit and use method thereof

Examples

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Embodiment 1

[0030] Embodiment 1: In this embodiment, the kit includes (1) Hep-2 cell antigen sheet; (2) fluorescently labeled secondary antibody; (3) fluorescence intensity calibration microspheres; (4) buffer; Tablet; (6) Instructions for experimental operation and quantitative analysis of fluorescence intensity. The Hep-2 cell antigen sheet was purchased from Germany Omen Medical Diagnostics Co., Ltd., and the fluorescently labeled secondary antibody was a Dylight488-labeled goat anti-human (IgG+H) antibody (maximum excitation wavelength: 493nm, maximum emission wavelength: 518nm ). The fluorescent intensity standard microspheres are made of polystyrene, with an average diameter of 5 μm (CV<3%) within a batch, uniform fluorescence intensity within a batch (CV<3%), a maximum excitation wavelength of 505 nm, and a maximum emission wavelength of 515 nm. The indirect immunofluorescence reaction procedure was carried out as follows:

[0031] 1. The first incubation: Add 25 μl of 1:100 dilu...

Embodiment 2

[0045] Embodiment 2: In this embodiment, the kit includes (1) He p-2 cell antigen sheet; (2) fluorescently labeled secondary antibody; (3) fluorescent intensity calibration microspheres; (4) buffer; (5) ) Mounting agent; (6) Instructions for experimental operation and quantitative analysis of fluorescence intensity. The Hep-2 cell antigen sheet was purchased from Oumen Medical Diagnostics Co., Ltd., Germany, and the fluorescently labeled secondary antibody was Alexa Fluor488-labeled goat anti-human (IgG+H) antibody (excitation wavelength: 493nm, emission wavelength: 518nm) . Fluorescence intensity standard microspheres are made of polystyrene, with an average diameter of 5 μm (CV<3%), uniform fluorescence intensity (CV<3%), a maximum excitation wavelength of 488nm, and a maximum emission wavelength of 510nm. The indirect immunofluorescence reaction procedure was carried out as follows:

[0046] 1. Add 25 μl of 1:100 diluted test serum to the prefabricated Hep-2 cell antigen ...

Embodiment 3

[0053] Embodiment 3: In this embodiment, the kit includes (1) Hep-2 cell antigen sheet; (2) fluorescently labeled secondary antibody; (3) fluorescent intensity calibration microspheres; (4) buffer; Tablet; (6) Instructions for experimental operation and quantitative analysis of fluorescence intensity. The Hep-2 cell antigen sheet was purchased from Oumen Medical Diagnostics Co., Ltd., Germany, and the fluorescently labeled secondary antibody was Alexa Fluor488-labeled goat anti-human (IgG+H) antibody (excitation wavelength: 493nm, emission wavelength: 518nm) . Fluorescence intensity standard microspheres are made of polystyrene cross-linked DVB, with an average diameter of 4 μm (CV<3%), uniform fluorescence intensity (CV<3%), a maximum excitation wavelength of 470nm, and a maximum emission wavelength of 510nm. The indirect immunofluorescence reaction procedure was carried out as follows:

[0054] 1. Add 25 μl of 1:100 diluted test serum to the prefabricated Hep-2 cell antige...

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Abstract

The present invention provides a kit for quantitative detection of homogeneous or speckled antinuclear antibody, and a use method thereof. The kit comprises: (1) a Hep-2 cell antigen sheet; (2) a fluorescence labeled secondary antibody; (3) fluorescence intensity calibration microspheres; (4) a buffer; (5) a sheet sealing agent; and (6) an experimental operation and quantitative fluorescence intensity analysis instruction. According to the present invention, the (1) Hep-2 cell antigen sheet provided by the kit, the (2) fluorescence labeled secondary antibody provided by the kit, and a serum sample to be detected are subjected to an indirect immunofluorescence reaction, the (3) fluorescence intensity calibration microspheres are added to the (1) Hep-2 cell antigen sheet to be detected, observing and microscopic image shooting are performed through a fluorescence microscope, a microscopic image analysis software is used to carry out fluorescence intensity analysis, and the fluorescence intensity value is converted into the corresponding serum antinuclear antibody titer value according to the (6) experimental operation and quantitative fluorescence intensity analysis instruction provided by the kit, such that the quantitative detection of the homogeneous or speckled antinuclear antibody in the serum sample is achieved.

Description

technical field [0001] The invention relates to a kit for quantitatively detecting antinuclear antibodies in human serum and a use method thereof. The kit is used for quantitatively detecting homogeneous or spot-type antinuclear antibodies in human serum. Background technique [0002] Antinuclear antibody (antinuclear antibody, ANA), also known as anti-nucleic acid antigen antibody, is a group of various components of eukaryotic cells, such as deoxyribonucleoprotein (DNP), DNA, extractable nuclear antigen (ENA) and RNA, etc. The general term for autoantibodies to target antigens, mainly present in serum. Antinuclear antibodies have varying degrees of positive rates in a variety of autoimmune diseases, such as systemic lupus erythematosus (SLE, 95% to 100%), rheumatoid arthritis (RA, 10% to 20%), mixed Connective tissue disease (MCTD, 80%-100%), Sjogren's syndrome (SjS, 10%-40%), systemic scleroderma (85%-90%), lupus hepatitis (95%-100%), Primary biliary cirrhosis (95% to 1...

Claims

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Application Information

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IPC IPC(8): G01N33/533
CPCG01N33/564G01N21/6428
Inventor 刘瑛颖王晶隋志伟张玲
Owner NAT INST OF METROLOGY CHINA
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