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Candida albicans attenuated live vaccine as well as preparation method and application thereof

A technology of Candida albicans and live bacteria, applied in the field of biomedicine, can solve the problems of high mortality rate of deep fungal infection, increase of fungal infection rate, loss of body flora, etc., and achieve good protection effect, short cycle and low cost Effect

Inactive Publication Date: 2013-08-21
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Most of the fungi are conditional pathogenic bacteria. The occurrence of clinical AIDS, the development of aggressive treatment methods, and the long-term unreasonable use of broad-spectrum antibiotics, adrenocortical hormones, cytotoxic drugs, and immunosuppressants have led to the loss of the body’s flora, resistance Decreased force, which greatly increases the incidence of fungal infection, and the fatality rate of deep fungal infection is extremely high

Method used

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  • Candida albicans attenuated live vaccine as well as preparation method and application thereof
  • Candida albicans attenuated live vaccine as well as preparation method and application thereof
  • Candida albicans attenuated live vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Deletion bacteria gpi7 / gpi7 The build:

[0033](1) Materials: Genomic DNA extraction kits, plasmid extraction kits, and DNA purification kits were purchased from Beijing Tiangen Gene Technology Co., Ltd. rTaq, exTaq, dNTP, etc. were purchased from Takara Company. The 96wells PCR instrument was purchased from BIO-RAD Company.

[0034] (2) Preparation of experimental reagents:

[0035] 1. PBS buffer

[0036] NaCl 8.0g, KCl 0.4g, NaCl 2 HPO 4 12H 2 O 3.48g, KH 2 PO 4 0.2g, adjust the pH to 7.4, add triple distilled water to 1000ml, dissolve and dispense, autoclave and store at 4°C for later use.

[0037] 2. Sandcastle Dextrose Agar Solid Medium (SDA)

[0038] Add 10g of peptone, 40g of glucose, and 18g of agar, add 900ml of triple-distilled water and stir to dissolve, dilute to 1000ml, autoclave (121°C, 15min) and store at 4°C for later use.

[0039] 3. Synthetic complete medium (synthetic complete medium, SC)

[0040] Yeast nitrogen source 6.7g, ...

Embodiment 2

[0088] Example 2. Virulence detection of deletion bacteria gpi7 / gpi7

[0089] Experimental animals: 6-8 week old C57 female mice, from Shanghai Slack Company. Certificate number: SCXK-(military) 2009-0004.

[0090] Experimental method and results:

[0091] C57BL / c female mice were equally divided into two groups, 12 in each group. Injected into tail vein of mice respectively gpi7 / gpi7 Deletion bacteria and parent bacteria SN152 each 5 × 10 5 CFU / only, causing systemic fungal infection. 15 rats (5 rats / group) were sacrificed 24 hours after the administration of bacteria, and both kidneys were taken, weighed, cut open, homogenized with 5ml of 0.9% normal saline, and then diluted with 0.9% normal saline for 10 2 、10 3The number of homogenates obtained by diluting multiples of different orders of magnitude was obtained, and 100ul each was spread on an SDA plate, cultured in a fungal incubator at 30°C for 48 hours, and the number of colonies was counted, with the number of ...

Embodiment 3

[0093] Example 3. Observation of deleted bacteria under electron microscope gpi7 / gpi7 cell wall:

[0094] Take a small amount of each strain from the glycerol frozen bacteria at -80°C, and inoculate it into YPD solid culture based on 30°C for 48 hours; pick a single colony from it and shake it in 1ml YPD liquid medium for 16 hours to activate it, so that the fungus is in the index Late growth period. Take 1ml of each of the above bacterial solutions in a 250ml Erlenmeyer flask, add 100ml of YEPD culture solution, incubate at 30°C and 200rpm for 14h, centrifuge at 1000×g for 10min, wash with PBS 3 times until the supernatant is colorless, remove the supernatant, add 3 % Glutaraldehyde 4ml was fixed overnight at 4°C. After washing with PBS for 3 times, it was fixed with osmic acid, dehydrated with ethanol and acetone gradient, and then mixed with acetone and embedding agent in different proportions to soak the sample. Finally, the sample was embedded, polymerized, positioned...

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Abstract

The invention relates to a candida albicans attenuated live vaccine for preventing candida albicans infection as well as a preparation method and application thereof. The vaccine is a candida albicans strain which contains a candida albicans GPI7 and is lack of a whole-genome function, so that candida albicans infection can be effectively prevented and candida albicans infection survival rate is effectively prolonged, and therefore, the candida albicans attenuated live vaccine can be used for preparing attenuated live vaccine for preventing the candida albicans infection.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a live attenuated Candida albicans vaccine, a preparation method and an application thereof. Background technique [0002] Most of the fungi are conditional pathogenic bacteria. The occurrence of clinical AIDS, the development of aggressive treatment methods, and the long-term unreasonable use of broad-spectrum antibiotics, adrenocortical hormones, cytotoxic drugs, and immunosuppressants have led to the loss of the body’s flora, resistance The force decreases, which greatly increases the occurrence of fungal infection, and the fatality rate of deep fungal infection is extremely high. Clinical statistical analysis at home and abroad found that most deep fungal infections in hospitals are caused by Candida, among which Candida albicans has the highest proportion. The body's resistance to Candida albicans infection is a complex process, including non-specific immunity and speci...

Claims

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Application Information

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IPC IPC(8): A61K39/02A61P31/04C12N15/87C12N7/01C12R1/01
Inventor 安毛毛陈思敏刘伟慎慧张石群黄鑫徐国彤姜远英
Owner TONGJI UNIV
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