Bird's nest colloidal gold immune test strip and preparation method thereof

A technology of colloidal gold and test strips, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low detection technology specificity, cumbersome detection operation, low sensitivity, etc. Simple, cost-effective detection

Inactive Publication Date: 2014-10-01
SHENZHEN ACAD OF METROLOGY & QUALITY INSPECTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] (1) PCR method: use specific gene fragments in bird's nest for PCR detection. This method has good specificity, but the detection time is long, requires expensive equipment and professional operators, and the finished product of bird's nest often requires thermal processing, which is easy to cause sample loss. DNA degradation in the medium, resulting in false negative test results;
[0005] (2) Electrophoresis method: analysis is based on the protein map of bird’s nest, but the ability to resist matrix interference is poor and the sensitivity is low. It is limited to the detection of bird’s nest and cannot be used for the detection of bird’s nest products;
[0006] (3) Specific component analysis method: use chromatography, spectroscopy and other means to identify the authenticity of various chemical components in bird’s nest, such as specific elements, amino acids, sialic acid, sugars, etc., but its specificity is not high , easily disturbed by artificial additives
[0007] Therefore, the existing detection technology has low specificity, is easily interfered by artificial additives, and has a narrow scope of application. The detection operation is cumbersome, the detection time is long, it needs to be equipped with large analytical instruments and requires professional operators, etc.

Method used

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  • Bird's nest colloidal gold immune test strip and preparation method thereof
  • Bird's nest colloidal gold immune test strip and preparation method thereof
  • Bird's nest colloidal gold immune test strip and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Separation and purification of target antigen.

[0034] The target antigen is bird's nest sialoglycoprotein, which consists of two proteins with the same antigenic determinant but different degrees of glycosylation modification, with molecular weights of 106 kDa and 128 kDa, respectively, and the isoelectric point ≤ 3.0.

[0035] The bird's nest was ultrasonically extracted with 7M urea for 10 minutes, centrifuged to take the supernatant, and separated and purified using a liquid phase focuser. The target protein was precipitated at the positive electrode and washed with deionized water to obtain the target protein. The target protein was verified by SDS-PAGE and 2-D PAGE, the results are as follows figure 1 As shown, no other foreign protein bands were seen, and the purity met the immune requirements.

Embodiment 2

[0036] Example 2 Preparation and purification of anti-sialoglycoprotein antibody.

[0037] Female balb / c mice were immunized with bird's nest sialoglycoprotein for 6 weeks, 3 in each group. For the first immunization injection, 100 μL of 100 μg / mL immune antigen was fully emulsified with the same amount of complete Freund’s adjuvant, and injected directly into the intraperitoneal cavity. After an interval of two weeks, take the sampled antigen, emulsify it with 100 μL of incomplete adjuvant, and inject it in the same way.

[0038] One day before cell fusion or on the same day, the Kunming rats were killed by pulling the neck, soaked in 70% alcohol, and disinfected the body surface; the Kunming rats were fixed on the wax board with pins, the abdomen was cut open on the ultra-clean workbench, and the peritoneum was picked up with small tweezers. Inject 5mL of RPMI-1640 complete culture solution (obtained by adding 15% volume percentage concentration of fetal bovine serum to G...

Embodiment 3

[0045] Example 3 Preparation of colloidal gold.

[0046] Take 1ml of 1% chloroauric acid solution, add 99ml of ultrapure water to form a final concentration of 0.01% chloroauric acid solution, after heating and boiling, take 1.6ml of 1% trisodium citrate and quickly add it to the boiling chloroauric acid solution at one time, Continue heating until the solution turns from light yellow to blue-black and finally bright red. After the color is stable, continue heating for 5 minutes, cool at room temperature, adjust pH to 5.9-6.2, and dilute to 100 mL with ultrapure water.

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Abstract

The invention provides a bird's nest colloidal gold immune test strip, comprising a sample pad, a binding pad, a reaction pad, a water-absorbing pad and a PVC backing, and the PVC backing is sequentially adhered to a sample pad, a binding pad, a reaction pad and a water-absorbing pad; the combination pad is adhered to the sample pad and overlapped with the reaction pad; the reaction pad is overlapped with the absorbent pad; the combination pad is coated with antisialosaccharide Protein antibody colloidal gold markers, the reaction pads are respectively coated with a detection line composed of sialoglycoprotein antigen and a quality control line composed of goat anti-mouse IgG. The invention can perform qualitative detection on the bird's nest and its related products, has wide application range, good specificity, good anti-matrix interference effect, can realize on-site rapid identification of the bird's nest and its related products, and has low detection cost.

Description

technical field [0001] The invention relates to the detection of bird's nest and related products, in particular to a bird's nest colloidal gold immune test strip and a preparation method thereof. Background technique [0002] Bird's nest is a nest formed by the condensation of saliva or down feathers of swiftlets of the Swiftlet family Swiftlet and similar species. Its origin is distributed in Southeast Asia, such as China, Indonesia, Thailand, Vietnam and Malaysia. Bird's nest is a kind of precious traditional Chinese medicine, which has the functions of nourishing yin and moistening dryness, nourishing the middle and nourishing qi, nourishing the stomach and spleen, resolving phlegm and relieving cough; bird's nest is rich in active glycoprotein, calcium, iron, phosphorus, iodine and vitamins and other natural nutrients And minerals, it also has the effect of preventing influenza virus, promoting cell growth and strengthening the function of the immune system. Due to the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558
Inventor 赖心田杨国武张世伟刘小青李芸严琼英陈血剑张秦蕾陈国培陈雁叶秀玲
Owner SHENZHEN ACAD OF METROLOGY & QUALITY INSPECTION
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