Candida chromogenic medium, detection kit and detection method

A technology of chromogenic medium and detection kit, which is applied in biochemical equipment and methods, measurement/inspection of microorganisms, etc., which can solve problems such as inconvenient, inability to identify other species, fluorescence cannot be directly observed by naked eyes, etc.

Active Publication Date: 2014-02-19
BEIJING JUNLIKANG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In terms of the culture medium for isolating and identifying Candida bacteria, Canadian patent CA2634325 describes the use of sweet potato extract powder and yeast extract powder as the main raw materials of the medium base, adding the chromogenic agent 5-bromo-4- Chloro-3-indolyl-N-acetyl-β-D-aminoglucopyranoside (5-Bromo-4-chloro-3-indolyl N-acetyl-β-D-glucopyranoside or X-N-acetyl-β- D-glucopyranoside) and the fluorescent substrate 4-methylumbelliferone-N-acetyl-β-D-galactosamine (4MU-N-acetyl-β-D-galactosaminide), by observing the colony color (blue , yellow and white) and whether the colonies produce fluorescence to distinguish the main species of Candida, but on the one hand, the fluorescence cannot be directly observed by the naked eye, which is inconvenient; on the other hand, light yellow and milky white are not easy to distinguish, which will lead to the bias in judgment
Japanese patent JP2003310298, describes a 5-bromo-4-chloro-3-indolyl-β-D-glucopyranoside (X-gluc) substrate to distinguish Candida albicans and Candida tropicalis from other fungi developed, but cannot identify other species of the genus
Venitia M. Cooke (New Chromogenic Agar Medium for the Identification of Candida spp., Applied and Environmental Microbiology, Vol.68 (7): 3622-3627, 2002) has described a kind of based on Sa Paul's culture medium and added assay Candida albicans ( C. albicans) and C. dubliniensis (C. dubliniensis) chromogenic substrate for glucosaminidase activity (VLPA-GlcNAc) to identify the chromogenic medium of Candida spp. White colonies with red dots, other Candida species can only be classified as white or pink colonies and cannot be distinguished
A. Gaschet et al. (Evaluation of CandiSelect4, a new chromogenic medium for isolation and presumptive identification of Candidaspecies from clinical specimens, J. of Medical Mycology, V.18(2), 89-95, 2008) described a new isolation and presumptive identification of CandiSelect4, a chromogenic medium for identifying Candida bacteria in clinical specimens, has the same separation effect as CHROMAgar Candida (Komagar Company) on Candida, but neither of them can completely obtain Candida albicans, which is important for this genus. Satisfactory identification results (a small number of false positive results) cannot distinguish Candida dublin from Candida albicans, and the target bacteria can be distinguished by the naked eye after at least 48 hours of culture, and the best time is 72 hours

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  • Candida chromogenic medium, detection kit and detection method
  • Candida chromogenic medium, detection kit and detection method
  • Candida chromogenic medium, detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Embodiment 1: Evaluation of nutritional capacity of Candida chromogenic medium according to the present invention

[0077] 1. Culture medium preparation

[0078] Weigh each component according to the formula in Table 1, prepare two culture media, add 1000mL of water to each, adjust the pH to 6.2 after heating and melting, boil for 2min, cool to 50°C, pour into a plate, and set aside.

[0079] Table 1 Medium composition (g / L)

[0080]

[0081] * The substrates for hexosaminidase and alkaline phosphatase are 5-bromo-4-chloro-3-indolyl-N-acetyl-β-D-glucosaminidase and 5-bromo-6-chloro-3- Indolyl Phosphate p-Toluidine Salt.

[0082] 2. Vaccination

[0083] Candida albicans (C.albicans, ATCC26278; 10231), Candida tropicalis (C.tropicalis, ATCC 750, 1369), Candida krusei (C.krusei, ATCC 6258, 34135), Candida glabrata (C. glabrata, ATCC 2001) was inoculated in yeast mold broth (Yeast MoldBroth, BD product, product number: 271120), cultured at 35°C for 24-48h, and then ...

Embodiment 2

[0087] Embodiment 2: Candida chromogenic medium application condition test of the present invention

[0088] 1 medium preparation

[0089] Weigh each component to prepare medium according to the formula in Table 2, add 1000mL of water, heat and melt, adjust the pH to 6.2, boil for 2min, cool to 50°C, pour into a plate, and set aside.

[0090] Table 3 Medium 3 components (g / L)

[0091]

[0092]

[0093] * The substrates for hexosaminidase and alkaline phosphatase are 5-bromo-4-chloro-3-indolyl-N-acetyl-β-D-glucosaminidase and 5-bromo-6-chloro-3- Indolyl Phosphate p-Toluidine Salt.

[0094] 2 inoculation

[0095] Candida albicans (C.albicans, ATCC26278; 10231), Candida tropicalis (C.tropicalis, ATCC 750, 1369), Candida krusei (C.krusei, ATCC 6258, 34135), Candida glabrata (C. glabrata, ATCC 2001) was inoculated in yeast mold broth (Yeast MoldBroth, BD product, product number: 271120), cultured at 35°C for 24-48h, and then used an inoculation loop to take a ring and inoc...

Embodiment 3

[0100] Embodiment 3: Sensitivity comparative test of Candida chromogenic culture medium and Sapaul culture medium according to the present invention

[0101] 1. Chromogenic medium plate preparation of the present invention

[0102] Take 3.0g of yeast powder, 3.0g of malt extract powder, 5.0g of peptone, 20.0g of glucose, 15g of agar, 0.05g of chloramphenicol, 2.0g of mixed chromogenic substrate, 1.0g of glycine, and 1000mL of water. After heating and melting, adjust the pH 6.4, boiled for 2 minutes, cooled to 50 ° C, poured plate, set aside.

[0103] 2. Control medium plate preparation

[0104] (1) Sapauer culture medium (BD Company of the United States, article number: 210950, hereinafter referred to as SDA)

[0105] Weigh 47g of dry powder (product of BD Company, serial number: 210950, 5.0g of caseytone, 5.0g of peptone, 20.0g of glucose, 17.0g of agar), add 1000mL of distilled water, pressurize at 121°C for 15min, cool to 50°C, invert the plate, and set aside.

[0106] (...

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Abstract

The invention relates to candida chromogenic medium, a detection kit and a detection method, belonging to the technical field of microbial diagnosis, in particular to the cultivation and identification of yeast in medical microbiology specimens, drugs and medical equipment sanitary inspection samples, public health surveillance samples and food (including cosmetics) sanitary inspection samples. The chromogenic medium of the invention is composed of basic medium, mixed chromogenic substrate and bacteriostat, wherein the mixed chromogenic substrate consists of aminocaproic glucosidase and alkaline phosphatase substrate, and candida specific enzyme is added into the mixed chromogenic substrate. The detection kit of the invention consists of the chromogenic medium, identification paper A containing enzyme substrate 5-bromo-4-chloro-3-indolyl-N-acetyl-beta-D-aminogalactose and identification paper B containing enzyme substrate 5-bromo-4-chloro-3-indolyl-beta-D- glycopyranoside. In the invention, the candida chromogenic medium and the detection kit have the advantages of low cost and simple configuration, and the method can be applied to the separation and identification of candida rapidly, simply and accurately.

Description

technical field [0001] The invention relates to the field of microbial diagnosis, especially the cultivation and identification of yeast in medical microbial specimens, hygienic inspection samples of medicines and medical devices, public health monitoring samples and food (including cosmetics) hygienic inspection samples, especially the isolation of Candida bacteria Culture and identification of important strains of the genus Candida albicans, Candida tropicalis, Candida krusei and Candida glabrata, and identification of Candida albicans. Background technique [0002] Candida (Candida spp., also known as Candida) is a kind of fungus, which not only widely exists in nature, but also usually exists in the oral cavity, upper respiratory tract, intestinal tract and vagina of normal people. Cause disease, when the immune function or general defense of the body is reduced or the mutual restriction of normal flora is out of balance, the bacteria multiply and change the growth form ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04
Inventor 赵贵明陈颖赵勇胜
Owner BEIJING JUNLIKANG BIOTECHNOLOGY CO LTD
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