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Sensitization detection method of colloidal gold immunity chromatography and use thereof

An immunochromatography, colloidal gold technology, applied in measurement devices, analytical materials, biological testing and other directions, can solve problems such as low sensitivity, and achieve the effect of improving sensitivity, improving detection sensitivity, wide application prospects and development value

Active Publication Date: 2009-07-01
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem solved by the present invention is to provide a sensitive detection method of colloidal gold immunochromatography. This method is based on the conventional colloidal gold immunochromatography assay method, using nano Gold particles process the detection line obtained by conventional colloidal gold immunochromatography to enhance the signal, which can significantly enhance the detection signal of conventional colloidal gold immunochromatography with clean background and low background value, which can improve the detection sensitivity by 8 to 200 times , thus overcoming the problem of low sensitivity of the traditional colloidal gold immunochromatographic detection method

Method used

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  • Sensitization detection method of colloidal gold immunity chromatography and use thereof
  • Sensitization detection method of colloidal gold immunity chromatography and use thereof
  • Sensitization detection method of colloidal gold immunity chromatography and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Preparation of various reagents

[0041] 1. Chlorauric acid aqueous solution: Weigh 1.0 g of chloroauric acid, dissolve it in 100 mL of double-distilled water to form a chloroauric acid aqueous solution with a concentration of 1% by weight, and shake well.

[0042] 2. Trisodium citrate aqueous solution: Weigh 1.0-1.5 g of trisodium citrate, dissolve it in 100 mL of double-distilled water to form an aqueous solution of trisodium citrate with a concentration of 1-1.5% by weight, and shake well.

[0043] 3. Phosphate buffered saline (PBS): Dissolve 8g sodium chloride (NaCl), 0.2g potassium chloride (KCl), 1.44g disodium hydrogen phosphate (NaCl) in 800mL distilled water 2 HPO 4 ) and 0.24g potassium dihydrogen phosphate (KH 2 PO 4 ), adjust the pH value of the solution to 7.4 with hydrochloric acid (HCl), add water to 1L, and shake well.

[0044] 4. Phosphate buffer (PB): 0.2g potassium chloride (KCl), 1.44g disodium hydrogen phosphate (Na 2 HPO 4 ) and ...

Embodiment 2

[0053] Example 2: Contrast Test of Sensitive Detection of Avian Influenza Virus NP Expression Protein

[0054] The NP protein of artificially expressed avian influenza virus was diluted 200 times with a developing agent, and then serially diluted until it was diluted to 200×2 8 Times, with the commercially available avian influenza virus general-purpose gold standard detection test strips (Anigen Company) to measure samples of each concentration, each sample was measured twice, and one strip was reserved for each batch of determination as a negative control. In the test, the test line and the control line show clear dark red, which means the sample is positive.

[0055] Using the sensitization detection method provided by the present invention, the sensitization reagent A (0.5% chloroauric acid) is mixed with the sensitization reagent B (10mM hydroxylamine hydrochloride) according to the volume ratio of 1:5 to make a signal amplification reagent, respectively Take 50 μL of ...

Embodiment 3

[0056] Example 3: Colloidal gold-enhanced detection test for H9N2 subtype avian influenza virus

[0057] Chicken embryos were inoculated with H9N2 subtype avian influenza virus, and the allantoic fluid of the harvested chicken embryos was serially diluted with a developing agent until it was diluted to 2 12 Allantoic fluid samples of various dilutions were measured with commercially available avian influenza virus general-purpose gold standard detection test strips (Anigen Company), each sample was measured twice, and one strip was reserved for each batch of determination as a negative control. In the test, the test line and the control line show clear dark red, which means the sample is positive.

[0058] Using the sensitization detection method provided by the present invention, the sensitization reagent A (1% chloroauric acid) is mixed with the sensitization reagent B (10mM hydroxylamine hydrochloride) according to the volume ratio of 1:2 to make a signal amplification re...

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Abstract

A sensitivity raising detection method of colloid gold immunity chromatography method comprises: when using general colloid gold immunity chromatography to check object sample, adding an object sample, adding signal amplifying agents of 10 to 80ul between the gold label antibody layer and the detection line; processing the detection line obtained by general colloid gold immunity chromatography method; reacting under room temperature for 1 to 10min; if the object sample contains antigen, the detection carrier is formed with a detection line and a control line as clear black or grey black color, to represent positive property, or if the object sample not contains antigens, the control line is black or grey black color, and the detection line has not color, to represent negative property. The method can significantly improve the sensitivity of colloid gold immunity chromatography reagent kit on the detection of biological macromolecules, having fast color development, lower background interference compared with general silver stain method, wide application prospect and high development value.

Description

technical field [0001] The invention relates to a macromolecular substance detection method, in particular to a sensitivity enhancement method for gold standard immunochromatographic detection of biomacromolecular substances and a method for making a kit by using the same. Background technique [0002] At present, the simple, fast, highly selective and highly sensitive detection of biological macromolecules has always been the direction of people's efforts. This is in the rapid diagnosis of diseases, health quarantine, animal and plant quarantine, food safety testing, environmental monitoring, etc. areas are important. Among them, the immunoassay (Immunoassay) based on the specific reaction of antigen and antibody has high selectivity and convenience, and its principle is simple, it can be produced and used without large-scale experimental equipment or large investment, and the average sample testing cost It is far lower than the instrumental analysis method and other chara...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/53
Inventor 邹明强李锦丰金涌薛强田世民齐小花
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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