Immune-colloidal gold test strip for detecting adulterant cow milk in camel milk and application thereof

A technology of colloidal gold test paper and camel milk, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of non-existence, expensive instrument operation, and high sensitivity

Pending Publication Date: 2019-09-03
XINJIANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection methods that can be applied to camel milk mixed with bovine milk mainly include: 1. Electrophoresis detection technology: Although this method is easy to operate, its sensitivity is low, and it is not suitable for the detection of a large number of samples
The second is liquid chromatography analysis: this method has high sensitivity and requires a small amount of sample for detection, which is suitable for the detection of a large number of samples, but this method requires expensive instruments and skilled instrument operators and the operation is complicated, so it is not suitable for on-site inspection
The third is nucleic acid differential analysis: this method has high sensitivity, but the extraction process of genetic material is relatively complicated
The fourth is immunological detection: Immunological detection is a method based on the specific reaction of antigen-antibody. The most widely used detection techniques are enzyme-linked immunosorbent assay and colloidal gold immunochromatography technology. The reaction results can be monitored by monitoring whether the reaction is There is a color change and the depth of the color change to distinguish whether there is a corresponding antigen or antibody in the test object, and it can qualitatively or quantitatively detect the target substance in the test object. However, in the existing detection methods, most of them use precision instruments for detection. There is no rapid and convenient colloidal gold immunochromatographic test strip technology for the detection of camel milk mixed with bovine milk

Method used

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  • Immune-colloidal gold test strip for detecting adulterant cow milk in camel milk and application thereof
  • Immune-colloidal gold test strip for detecting adulterant cow milk in camel milk and application thereof
  • Immune-colloidal gold test strip for detecting adulterant cow milk in camel milk and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1: SDS-PAGE compares the protein composition of cow milk and camel milk

[0061] Camel milk whey, bovine milk whey, camel milk casein, and bovine milk casein were subjected to polyacrylamide gel electrophoresis (SDS-PAGE), Coomassie brilliant blue stained gel and decolorized to record the results with a gel imager, see attached figure 1 As shown, it was found that camel whey lacked β-lactoglobulin relative to bovine whey, and the protein content of α-lactalbumin in camel whey was significantly higher than that in bovine whey. Therefore, bovine milk β-lactoglobulin and camel milk α-lactalbumin were selected for Preparation of polyclonal antibody; Camel milk casein contains less κ-casein and α-casein than bovine milk s1 - Casein has a large molecular weight, so cow's milk κ casein and camel milk α are selected s1 - Casein for the preparation of polyclonal or monoclonal antibodies.

Embodiment 2

[0062] Example 2: Camel milk α-lactalbumin and α s1 Extraction of casein

[0063] (1) Crude extraction of camel milk α-lactalbumin by ammonium sulfate fractional precipitation

[0064] Take defatted camel milk, adjust the pH to 4.3-4.8, centrifuge at 3000-12000r / min for 10-60min, and repeat the centrifugation several times. The supernatant is subjected to fractional precipitation according to the ammonium sulfate saturation of 70%-100%, and the obtained precipitate is desalted and freeze-dried spare.

[0065] (2) Calcium ion precipitation crude extraction of camel milk α s1 -casein

[0066] Take defatted camel milk, adjust the pH to 10-12, add calcium ions to produce a precipitate, adjust the pH to 6-8, centrifuge at 3000-8000r / min for 10-30min, take the precipitate and dissolve it in 3.3mol / L urea, adjust the pH to 4 ~5, centrifuge at 3000~8000r / min for 10~30min, get the precipitate and dry it for later use.

[0067] (3) Ultrafiltration and centrifugation for coarse sepa...

Embodiment 3

[0071] Example 3: Cow milk β-lactoglobulin, cow milk κ-casein, camel milk α-lactalbumin, camel milk α s1 - Preparation of monoclonal antibody against casein

[0072] (1) Bovine milk β-lactoglobulin, bovine milk κ-casein, camel milk α-lactalbumin, camel milk α s1 - Preparation of monoclonal antibody against casein:

[0073] Female Balb / c mice about 7 weeks old were selected as immunized animals, and the immunogens were bovine milk β-lactoglobulin, bovine milk κ-casein, camel milk α-lactalbumin and camel milk α s1 - Casein, immunogen and Freund's complete adjuvant (primary immunization) or Freund's incomplete adjuvant (boost immunization) immunization, after ultrasonic mixing and emulsification, multi-point subcutaneous and intraperitoneal injection to immunize mice. The titer of the isolated serum was determined by indirect ELISA method. When the titer of the serum reached above 1:64000, the mouse with the highest reaction value was injected into the tail vein and peritoneal ...

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Abstract

The invention discloses an immune-colloidal gold test strip for detecting adulterant cow milk in camel milk and application thereof. A camel milk alpha-lactalbumin gold-labeled antibody, a camel milkalphas1-casein gold-labeled antibody, a cow milk beta-lactoglobulin gold-labeled antibody and a cow milk k-casein gold-labeled antibody are prepared; a test strip I for detecting a whey protein component and a test strip II for detecting a casein component are prepared respectively; and then an immune-colloidal gold test strip for detecting adulterant cow milk in camel milk is obtained. The test strip is capable of monitoring the cow source and camel source protein of the milk and the dairy product simultaneously and has the high practicability and the great development value in the adulteration monitoring field of the camel milk.

Description

technical field [0001] The present invention mainly relates to the technical field of immune detection, in particular, the present invention relates to the technical field of colloidal gold detection for detecting adulterated cow milk in camel milk. Background technique [0002] With the development of the economy and the enhancement of people's awareness of health care, camel milk products have gradually entered people's daily life. Camel milk is rich in nutrients, and the content of immunoglobulin, lactoferrin, insulin and insulin-like growth factors in protein active ingredients is relatively high. The structure of camel milk α-lactalbumin is easier to absorb than cow milk α-lactalbumin, and camel milk does not contain β-lactoglobulin, the main allergen in cow milk, so it is more suitable for babies who are allergic to cow milk. The oligosaccharides, unsaturated fatty acids, and minerals in camel milk have special composition patterns and are easy to digest. In recent ye...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/31
CPCG01N33/68G01N33/558G01N33/553G01N33/531G01N33/532G01N21/31
Inventor 杨洁邓露李敏婧王雪岳海涛申彤陈红征
Owner XINJIANG UNIVERSITY
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