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Method for indicating xenotropic antibody interference in immune reaction

A heterophilic antibody and heterophilic technology, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as misdiagnosis and false negative rate increase

Active Publication Date: 2007-01-03
SHANGHAI TELLGEN LIFE SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the existence of the HD-HOOK effect, the detected sample cannot be correctly distinguished whether its concentration exceeds the linear range of the detection kit or the concentration itself is the value, so that the experiment is misdiagnosed, especially resulting in an increase in the false negative rate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0145] Parallel detection of 10 tumor markers (no HID-HOOK indication, no heterophile antibody interference indication)

[0146] 1. Prepare

[0147] 1.1 The primary antibody removes amino-containing small molecules and foreign proteins (dialysis or chromatography column) in advance, and measures their concentration.

[0148] 1.2 In a 1.5ml polypropylene centrifuge tube, accurately weigh about 5mg of N-hydroxysulfosuccinimide (NHS), and set aside (moisture-proof).

[0149] 1.3 In a 1.5ml polypropylene centrifuge tube, accurately weigh about 5mg of N-ethyl-N'(3-dimethylainopropyl)-carbodiimide (EDC) for future use (moisture-proof).

[0150] 2. Beads activation

[0151] 2.1 Suspend the microsphere stock solution in a vortex mixer for 20 seconds, pipette 200 μL of microspheres (equivalent to 2.5×10 6 microspheres) in a 1.5ml polypropylene centrifuge tube.

[0152] 2.2 Centrifuge at 15000rpm for 2 minutes (set for 3 minutes), remove the supernatant.

[0153] 2.3 Add 100 μL of ...

Embodiment 2

[0190] Indication of HAMA reaction in human serum samples and detection of alpha-fetoprotein (AFP) in serum

[0191] Antibodies against HAMA were purified mouse IgG purchased from Biodesign, primary and secondary antibodies against AFP were purchased from Shanghai Second Military Medical University, AFP standard products were purchased from Biodesign, and Beads were purchased from Luminex.

[0192] 1. Prepare

[0193] 1.1 The primary antibody and AFP antigen were removed in advance of amino acid-containing small molecules and foreign proteins (dialysis or chromatographic column), and their concentrations were measured.

[0194] 1.2 In a 1.5ml polypropylene centrifuge tube, accurately weigh about 5mg of N-hydroxysulfosuccini-mide (NHS), and set aside (moisture-proof).

[0195] 1.3 In a 1.5ml polypropylene centrifuge tube, accurately weigh about 5mg of N-(3-dimethylainopropyl)-N-ethyl-carbodiimide (EDC) for future use (moisture-proof).

[0196] 2. Microsphere (beads) activatio...

Embodiment 3

[0239] Parallel detection of 5 tumor markers (with HAMA indication)

[0240] Basically according to the steps of Examples 1 and 2, the parallel detection of five tumor markers with HD-HOOK indication was carried out, with the difference that SCCA, CA125, CA15-3, CEA, and β-HCG were selected as the antigens to be detected. One detection bead and HAMA indicator bead were used for each, and the 5 detection bead and 5 indicator bead had different coded fluorophores.

[0241]

[0242] Use pH 7.4 phosphate buffer to prepare solution B according to the above table.

[0243] In addition, the preparation of liquid A, liquid C and liquid H is the same as in Examples 1 and 2.

[0244] SCCA

[0245] Judgment of the experimental results: samples 1-5 are HAMA-negative samples, and samples 6-10 are HAMA-positive samples.

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PUM

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Abstract

The invention is concerned with the out-body examination field that indicates the interferential method and the corresponding reagent box of the Heterophilic antibody. The invention can improve the accuracy of the double-position filling immunoassay, and reduce the false positive ratio of the double-position filling immunoassay.

Description

technical field [0001] The invention relates to the technical field of in vitro detection, in particular to a method for indicating the interference of heterophilic antibodies in the immunological detection of serum samples. Background technique [0002] Heterophilic antibody interference refers to the protein antibody substance that directly connects the capture antibody and the detection antibody without forming a sandwich in the immune double-antibody sandwich reaction to form a false positive result. Common heterophilic antibodies include HAMA (Human Anti-mouse Antibody) antibody and rheumatoid factor. [0003] A common heterophilic antibody interference is caused by HAMA reactions. HAMA (Human Anti-mouse Antibody) reaction often occurs in immunoassays, and its incidence is about 10%. For example, human anti-mouse antibodies will be present in individuals who have been exposed to or vaccinated against murine antibodies. Since the serum samples from these individuals c...

Claims

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Application Information

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IPC IPC(8): G01N33/538G01N33/543
Inventor 姚见儿周雪雷罗朝领茅柳娟
Owner SHANGHAI TELLGEN LIFE SCI CO LTD