Colloidal gold test strip for detecting hereditary hearing loss, and preparation method and application thereof

A technology for hereditary deafness and colloidal gold test paper, which is applied in the field of colloidal gold to achieve the effects of accurate results, simple operation and low cost

Inactive Publication Date: 2016-04-27
WUHAN J H BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Gold standard immunochromatographic technology has not yet be

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  • Colloidal gold test strip for detecting hereditary hearing loss, and preparation method and application thereof

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preparation example Construction

[0026] The preparation method of the above-mentioned colloidal gold test paper card for detecting hereditary deafness comprises:

[0027] (1) preparation of colloidal gold solution: in HauCl Add trisodium citrate, after boiling, use trisodium citrate, tannic acid as reducing agent to obtain the colloidal gold solution that diameter is 10-20nm;

[0028] (2) Preparation of colloidal gold binding pad: adjust the colloidal gold solution to pH 7.5-8.5 and add SLC26A4 antibody, GJB2 antibody and blocking agent. After labeling, redissolve the precipitate into the colloidal gold solution containing BSA and sucrose. The reconstituted product is added to glass fiber membrane or polyester membrane and then dried;

[0029] (3) Chromatography pad preparation: SLC26A4 antibody and GJB2 antibody were diluted with PBS, respectively streaked on the nitrocellulose membrane and dried;

[0030] (4) Test paper card assembly: Paste the sample pad, colloidal gold binding pad, chromatographic membra...

Embodiment 1

[0034] This embodiment provides a colloidal gold test card for detecting hereditary deafness, and provides a preparation method of the test card.

[0035] 1. Main materials

[0036] SLC26A4, GJB2 specific paired antibody, goat anti-mouse IgG antibody: Beijing Shengqi Yanghuan Technology Co., Ltd.; chloroauric acid: Sigma company; nitrocellulose (NC) membrane: SARTORIUS (Germany), CN140; Germany Sartorius company Products; bovine serum albumin (BSA), polyethylene glycol PEG20000, hydrolyzed casein: Sigma products. Other commonly used reagents are analytical reagents.

[0037] Two, the preparation method of colloidal gold

[0038] 1. Add HauCl 4 Prepare 0.01% aqueous solution first, take 100ml and heat to boiling.

[0039] 2. Accurately add 1% trisodium citrate (Na 3 C 6 h 5 o 7 2H 2 O) 2.00-4.50ml of aqueous solution.

[0040] 3. Continue heating and boiling for 30 minutes. At this time, it can be observed that the light yellow chloroauric acid aqueous solution turns...

Embodiment 2

[0058] This embodiment provides a colloidal gold kit for detecting hereditary deafness.

[0059] The test kit includes a test paper, a colorimetric card or a colloidal gold quantitative reader, and a plastic card for accommodating the test paper. The test paper is loaded into the plastic card to form a test paper card. Specifically, the test kit also includes a dropper Tubes, Serum Cups and Disposable Syringes. The detection test paper is the colloidal gold test paper card provided in Example 1 for detecting hereditary deafness. The colloidal gold quantitative reader is a known optical detection system.

[0060] The samples applicable to this kit are the same as those applicable to the colloidal gold test paper card for detecting hereditary deafness. Serum, whole blood, urine, etc. can be used as detection samples. In addition, tissue sections, cell smears and other samples for histochemical qualitative tests Can be used as samples.

[0061] The above kit was used to detect...

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Abstract

The invention provides a colloidal gold test strip for detecting hereditary hearing loss. The colloidal gold test strip comprises a base plate, and a sample loading pad, a colloidal gold binding pad, a chromatographic membrane and a sample sucking pad which are successively adhered on the base plate, wherein the colloidal gold binding pad is a glass fiber membrane or polyester film coated with colloidal gold-labeled SLC26A4 antibody and GJB2 antibody and the chromatographic membrane is a cellulose nitrate membrane coated with SLC26A4 and GJB2 antibodies. The invention further provides a preparation method for the colloidal gold test strip and a colloidal gold kit for detecting hereditary hearing loss. According to the invention, colloidal gold immunochromatography is employed to simultaneously detect the SLC26A4 and GJB2 antibodies; expensive apparatuses and tedious operation are not needed in detection; detection flow is simplified; a detection period is substantially shortened; people can detect hereditary hearing loss by themselves in virtue of the colloidal gold test strip and kit; whether gene mutation exists in a to-be-tested sample can be confirmed, so reference is provided for clinical diagnosis and treatment; and the colloidal gold test strip and kit are extensively applied to large-scale pre-pregnant prevention, intervention in pregnancy, postpartum screening and medication directions.

Description

technical field [0001] The invention belongs to the technical field of colloidal gold, and in particular relates to a colloidal gold test paper card for detecting hereditary deafness with a short detection period, a simple process and accurate results, a preparation method and an application thereof. Background technique [0002] Deafness is one of the most common disabling diseases and clinical genetic diseases. The reported incidence of newborns reaches 1-3‰. Studies have found that about 60% of newborns' deafness is related to genetic factors, that is, hereditary deafness. Hereditary deafness is divided into nonsyndromic hearing impairment (NSHI) and syndromic hearing impairment (SHI). About 70% of them are inherited in an autosomal recessive manner. Hereditary deafness is caused by mutations in a single gene or compound mutations in different genes. More than 120 deafness-related genes have been discovered so far, with high genetic heterogeneity of genes and loci. [...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/543
CPCG01N33/558G01N33/54306G01N2800/14
Inventor 胡军瑜王健斌龚贻洲周军
Owner WUHAN J H BIO TECH
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