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Reagent card for accurately detecting test object, kit and application

A test substance and reagent card technology, applied in the biological field, can solve the problems of failure to improve the impact, increase the probability of false negatives, misdiagnosis, etc., to avoid the hook effect, reduce the false negative rate, and improve the accuracy. Effect

Inactive Publication Date: 2016-10-26
XIAMEN BIOTIME BIOTECHNOLOGY CO LTD
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  • Summary
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AI Technical Summary

Problems solved by technology

The existence of this effect makes it impossible for the tested sample to correctly distinguish whether its concentration exceeds the linear range of the detection kit or the concentration itself is the value that causes this phenomenon, so that the experiment is misdiagnosed, especially resulting in an increase in the probability of false negatives
At present, there is no effective method to avoid the influence of the HOOK effect. Most of them improve the situation by increasing the amount of coated protein, increasing the amount of labeled protein or gold spray, or playing an auxiliary role through the adjustment of the process formula, but all Failed to improve the impact of this effect

Method used

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  • Reagent card for accurately detecting test object, kit and application
  • Reagent card for accurately detecting test object, kit and application
  • Reagent card for accurately detecting test object, kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: Preparation and detection of HCG detection kit (fluorescence immunochromatography)

[0025] Preparation of HCG antibody 1-coupled fluorescent microspheres and biotin-coupled fluorescent microspheres: select 10 mg of fluorescent microspheres and add them to 1 mL of 0.01M pH7.4 PBS solution containing 5% glutaraldehyde to react for 1 hour, and centrifuge at 12,000 rpm for 5 minutes. After removing the supernatant, add 1 mg of HCG Antibody 1 and mix evenly, react at 4°C for 12 hours, centrifuge at 12,000 rpm for 5 minutes, remove the supernatant, and resuspend with 1 mL of 0.01M pH7.4 PBS to form HCG Antibody 1-coupled fluorescent microspheres; use the same method 10 mg of fluorescent microspheres were selected and added to 1 mL of 0.01 M pH7.4 PBS solution containing 5 mass % glutaraldehyde for 1 h, centrifuged at 12,000 rpm for 5 min to remove the supernatant, then added 1 mg of biotin to mix, reacted at 4°C for 12 h, 12,000 rpm After centrifuging for 5 min...

Embodiment 2

[0054] Embodiment 2: AFP detection kit (fluorescence immunochromatography)

[0055] Compared with Example 1, the difference is that HCG antibody 2 is replaced by AFP antibody 2, and HCG antigen is secretly replaced by AFP antigen, and other methods and steps are the same as in Example 1.

[0056] Table 2 The detection result table of the standard product

[0057]

[0058]

[0059] From Figure 4 It can be seen that the logarithmic concentration-logarithmic T1 fluorescence of AFP concentration in the range of 5-400ng / mL presents a good linear relationship, Figure 5 In the range of AFP concentration in the range of 400-2000ng / mL, the logarithmic concentration-logarithmic T2 fluorescence presents a good linear relationship, and the concentration greater than 2000ng / mL does not appear HOOK effect. Table 2 shows that the cut-off point is 4.186.

[0060] Precision test: intra-batch precision, take the same batch of test cards and buffer solution, use the same known concent...

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Abstract

The invention discloses a reagent card for accurately detecting a test object, a kit and application. A sample pad (1), an antibody carrier film (2) and water absorption paper (3) are successively adhered to a PVC bottom plate (6) from one end to the other end, wherein the sample pad is a sample loading area, and the sample pad is sprayed with a test object antibody-1 coupling marker and a biotin coupling marker; the antibody carrier film is provided with three lines which are successively a line T1 (4), a line T2 and a line C (5) from the sample pad to the water absorption paper, or are the line T1 (4), the line C (5) and the line T2; the line T1 is a detection line, and a test object antibody 2 is smeared on the position of the line T1; an antigen of the test object is smeared on the position of the line T2; streptavidin is smeared on the position of the line C. By adopting the reagent card, the false negative rate caused by the high-dose hook effect can be greatly reduced, the clinical detection accuracy is improved, and the adverse impact of the hook can be effectively solved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a reagent card, kit and use for solving the high-dose hook effect. Background technique [0002] In clinical immunochromatographic experiments, when there is a high dose, it is easy to have a high-dose hook effect, that is, the HD-HOOK effect. It refers to the high-dose section of the dose-response curve in the two-site sandwich immunoassay. The linear trend is not flat and infinitely delayed, but curved downward, like a hook or a sickle. According to this Phenomenon, called "HD-HOOK" effect. The existence of this effect makes it impossible for the tested sample to correctly distinguish whether its concentration exceeds the linear range of the detection kit or the concentration itself is the value that causes this phenomenon, so that the experiment is misdiagnosed, especially resulting in an increase in the probability of false negatives. At present, there is no effective method to...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/543
CPCG01N33/558G01N33/54313
Inventor 颜珊洪国粦吴颖杨莉莉
Owner XIAMEN BIOTIME BIOTECHNOLOGY CO LTD
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