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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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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 there are traditional pathogenic microbiological isolation and identification, PCR and TCTA color detection methods for the diagnosis of the bacteria, these methods require high technical requirements, and are time-consuming and uneconomical.

Method used

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  • Rapid diagnosis test strip of cow mastitis candida albicans

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation of Candida albicans polyclonal antibody

[0027] 1 Materials and methods

[0028] 1.1 Strains: standard Candida albicans, clinical identification of Candida albicans, standard strain of Staphylococcus aureus, standard strain 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 and 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 methods

[0032] 2.1 Antigen preparation of Candida albicans

[0033] Streak the preserved isolated and identified Candida albicans on the Sabouraud solid medium, culture in a 37°C incubator for 18-48 hours, pick a si...

Embodiment 2

[0039] Embodiment 2 Preparation of Candida albicans monoclonal antibody

[0040] 1 Materials and methods

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

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

[0043] 2. Experimental method

[0044] 2.1 Immunization of experimental animals

[0045] Six Balb / c female mice aged 6-7 weeks were selected for immunization. Primary immunization, 0.25mL2×10 7 cfu mL -1 The inactivated Candida albicans was mixed with complete Freund's adjuvant in equal proportions, phacoemulsified, and injected subcutaneously at multiple points in the abdomen. After 3 weeks, the second immunization, 0.25mL2×10 7 cfu mL -1 Inactivated Candida albicans was mixed with incomplete Freund's adjuvant in equal proportions, and injected subcutane...

Embodiment 3

[0068] The preparation of embodiment 3 colloidal gold body

[0069] 1. Materials and methods

[0070] 1.1 Main reagent: HAuCl 4 , rabbit anti-Candida albicans polyclonal antibody, 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, air 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 trisodium citrate reduction method, with 500mL beaker, add 2.4mL1% HAuCl 4 In 240mL of distilled water (final concentration: 0.01%), divide into six 80mL beakers, each 40mL. Heat the induction cooker to boiling, add 1% trisodium citrate, each 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1mL, 1.2mL (marked as: 1, 2, 3, 4, 5, 6 respectively) , continue heating and stirr...

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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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