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Hepatitis c virus antibody time-resolved fluoroimmunoassay and kit

A hepatitis C virus, time-resolved technology, applied to the analysis of materials, measuring devices, instruments, etc., can solve the problems of poor stability, cumbersome operation, and low sensitivity, and achieve small cross-reaction, short detection time, and good specificity Effect

Inactive Publication Date: 2011-11-09
PERKINELMER MEDICAL DIAGNOSTICS PROD SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This technology has several benefits that make it better than previous methods for detecting hepatitis C virus. It can be highly sensitive but quickly detected within 6 hours without being affected by other factors like blood clotting. Additionally, this method works well even if there've been any new ones discovered during testing.

Problems solved by technology

This patented technical problem addressed in this patent relates to improving the accuracy and reliability of detecting viruses like Human Immunodeficiency Virustase (HAVi)-associated Non Anon B Hepatic Failure Syndrome ( NAFBS); specifically for identifying individuals who may be at risk due to poor health outcomes associated with these diseases.

Method used

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  • Hepatitis c virus antibody time-resolved fluoroimmunoassay and kit
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  • Hepatitis c virus antibody time-resolved fluoroimmunoassay and kit

Examples

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preparation example Construction

[0025] Next is the preparation of HCV biotin-labeled antigen, including the following steps:

[0026] (1) Add the antigen to be labeled into the dialysis bag, put it into the prepared pH 9.5 carbonate buffer, and dialyze overnight at room temperature (pH 9.5 carbonate buffer);

[0027] (2) The next day, take out the antigen, dilute the dialyzed antigen with pH 9.5 carbonate buffer to 0.2-5 mg / mL, and add biotin while shaking at a mass ratio of 2:1 (biotin: antigen). At room temperature, vibrate slowly on a vibrating plate machine for 1 hour, then dialyze overnight in phosphate buffered saline (pH7.5 phosphate buffer) after removal;

[0028] (5) filter with a filter with an aperture of 0.2um;

[0029] (6) Add 0.3% (W / V) premium pure BSA and store at 2-8°C.

Embodiment

[0030] Example: Hepatitis C virus antibody detection

[0031] 1. Biotin-labeled antigen

[0032] Dialyze the HCV recombinant antigen with carbonate buffer solution overnight, then add biotin to react for 1 hour, take it out and dialyze overnight in phosphate buffer solution with pH 7.5. Aliquot and store at +2~+8℃.

[0033] 2. Operation steps

[0034] 1.) Kit preparation: antigen-coated buffer (carbonate buffer), blocking solution (phosphate buffer and 2% (W / V) BSA), reaction buffer (50mmol / L of pH 7.8 Tris-HCl, containing 0.9% NaCl, 1% BSA, 0.5% casein, 0.08% Tween20 and 0.1% NaN 3 ), working washing solution (phosphate buffer saline), fluorescence enhancing solution (β-diketone body), as streptavidin coated with HCV recombinant antigen, biotin-labeled antigen and lanthanide ion-labeled.

[0035] A: Working washing solution: dilute 40mL phosphate buffer solution (25 times concentrate) with distilled or deionized water to 1,000mL for later use.

[0036] B: Biotin-labeled ...

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Abstract

The invention discloses a hepatitis c virus antibody time-resolved fluoroimmunoassay and a kit; a multi-fragment HCV fusion recombinant antigen is selected as a coating antigen, and a sodium carbonate buffer solution diluted into 1-10 mug/mL is used as a coating solution; a matched HCV recombinant antigen labelled with biotin is used as a labelled antigen; streptavidin is marked with lanthanum series elements; during experiment, a reaction buffer solution is diluted according to a ratio of 1:21 for using; on a reaction plate with the coating antigen, 25-muL HCV negative and positive controls or samples to be tested, and the diluted biotin-labelled antigen are added orderly into each hole; after incubation and plate washing, the diluted streptavidin europium marker is added; and fluorescence detection is performed after incubation. The invention also provides a corresponding kit. The invention has high sensitivity, specificity and stability; and the analysis system has high automation, can increase the speed for clinical detection results, can greatly reduce artificial error and improve the reliability of the detected results.

Description

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Claims

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Application Information

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Owner PERKINELMER MEDICAL DIAGNOSTICS PROD SHANGHAI
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