Rapid diagnosis test strip of cow mastitis candida albicans

A technology for Candida albicans and cow mastitis, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of uneconomical, intractable mastitis, time-consuming, etc., and achieve good stability, good specificity, and high sensitivity Effect

Inactive Publication Date: 2014-03-05
GUANGXI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This bacterium invades the mammary gland and reproduces in the mammary gland, causing frequent occurrences of mastitis in dairy cows, especially refractory mastitis. economic loss; at the same time, it also threatens human health
Although t

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  • Rapid diagnosis test strip of cow mastitis candida albicans

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Example Embodiment

[0026] Example 1 Preparation of Candida albicans polyclonal antibody

[0027] 1Materials and methods

[0028] 1.1 Strains: standard Candida albicans, clinically identified Candida albicans, standard strains of Staphylococcus aureus, standard strains of Escherichia coli.

[0029] 1.2 Main reagents and instruments: Martin broth, Sabouraud nutrient agar, horseradish peroxidase (HRP) labeled goat anti-rabbit IgG, Freund's complete adjuvant / Freund's incomplete adjuvant, dialysis bag, electrophoresis apparatus, Electrophoresis tank, UV gel imaging system, nucleic acid protein analyzer, ultra-high-speed refrigerated centrifuge.

[0030] 1.3 Experimental animals: New Zealand white rabbits (Animal Experiment Base of Guangxi University).

[0031] 2 Experimental method

[0032] 2.1 Antigen preparation of Candida albicans

[0033] The preserved, isolated and identified Candida albicans was streaked on Sabouraud solid medium, cultured in a 37°C incubator for 18 to 48 hours, and a single colony was pi...

Example Embodiment

[0039] Example 2 Preparation of Candida albicans monoclonal antibody

[0040] 1Materials and methods

[0041] 1.1 Experimental materials: Sp2 / 0 mouse myeloma cells, Candida albicans, Staphylococcus aureus standard strain, Escherichia coli standard strain, Streptococcus; cell culture dish, 50mL sharp-bottomed centrifuge tube, 96-well microtiter plate, dialysis bag Wait.

[0042] 1.2 Experimental animals: Balb / c mice.

[0043] 2. Experimental method

[0044] 2.1 Test animal immunity

[0045] Six Balb / c female mice aged 6-7 weeks were selected for immunization. Initial immunization, 0.25mL2×10 7 cfu·mL -1 Inactivated Candida albicans and Freund's complete adjuvant are mixed in equal proportions, phacoemulsified, and injected into multiple points under the skin of the abdomen. After 3 weeks, the second immunization, 0.25mL2×10 7 cfu·mL -1 Inactivated Candida albicans and Freund's incomplete adjuvant are mixed in equal proportions and injected subcutaneously at multiple points on the back....

Example Embodiment

[0068] Example 3 Preparation of colloidal gold body

[0069] 1. Materials and methods

[0070] 1.1 Main reagent: HAuCl 4 , Rabbit polyclonal antibody against Candida albicans, trisodium citrate, bovine serum albumin (BSA), skimmed milk powder, PEG20000, K 2 CO 3 .

[0071] 1.2 Main instruments: high-speed refrigerated centrifuge, electronic platform scale, gas bath constant temperature oscillator, DK-SD electric heating constant temperature water tank, ultraviolet-visible spectrophotometer.

[0072] 1.3 Preparation of colloidal gold by trisodium citrate reduction method

[0073] The preparation of colloidal gold adopts the trisodium citrate reduction method, using a 500mL beaker, adding 2.4mL 1% HAuCl 4 In 240mL of distilled water (final concentration of 0.01%), separate into 6 80mL beakers, 40mL each. Heat the induction cooker to boiling, add 1% trisodium citrate, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1mL, 1.2mL (respectively labeled: 1, 2, 3, 4, 5, 6) , Continue to heat and stir, observe the ...

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Abstract

The invention discloses a rapid diagnosis test strip of cow mastitis candida albicans. The test strip comprises an absorbent paper, an analysis membrane, a gold-labelled antibody joint pad, a sample pad and a base plate, wherein the analysis membrane contains a detection line and a quality control line; the detection line is enveloped by a candida abbicans monoclonal antibody; the quality control line is enveloped by goat anti-rabbit IgG; a colloidal gold-labelled anti-rabbit polyclonal antibody of the candida albicans is enveloped by the gold-labelled antibody joint pad. The rapid diagnosis test strip is applied to detection of cow mastitis pathogens, so that the rapid diagnosis test strip is specific, accurate, rapid, simple to operate and the like, and is applicable to basic livestock breeding, husbandry technology popularization, and animal husbandry and animal quarantine inspection. Therefore, the rapid diagnosis test strip has important value on rapid diagnosis of dairy cow mastitis.

Description

technical field [0001] The invention belongs to the technical field of dairy cow mastitis detection, in particular to a rapid diagnosis test strip for candida albicans of dairy cow mastitis. Background technique [0002] Candida albicans (Candida albicans), also known as Candida albicans, is one of the most common and major comorbid condition pathogenic fungi in human and animal fungal diseases. This bacterium invades the mammary glands and reproduces in the mammary glands, causing frequent occurrences of mastitis in dairy cows, especially refractory mastitis, which not only restricts the development of the dairy industry, but also seriously affects the quality of raw milk, causing great harm to the dairy farming industry. economic loss; at the same time, it also threatens human health. Although there are traditional pathogenic microbiological isolation and identification, PCR and TCTA color detection methods for the diagnosis of this bacteria, these methods require high te...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/558G01N33/531G01N33/56911
Inventor 刘平何宝祥黄云飞
Owner GUANGXI UNIV
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