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Fungus detection method based on liquid-based slide making technology and fluorescent staining technology

A fluorescent dyeing and film-making technology, which is applied in the field of fungal detection, can solve the problems of many reagents required for matching, troublesome storage and management, and high professional skills requirements, so as to achieve clear fungal morphology and structure, improve sensitivity and efficiency, and facilitate search and identification Effect

Inactive Publication Date: 2019-02-19
江西业力医疗器械有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Disadvantages: 1. The requirements for the professional skills of the operators are relatively high
2. The operation is cumbersome and takes a long time (about 1 hour) 3. There are many reagents required for matching, and storage management is troublesome

Method used

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  • Fungus detection method based on liquid-based slide making technology and fluorescent staining technology
  • Fungus detection method based on liquid-based slide making technology and fluorescent staining technology
  • Fungus detection method based on liquid-based slide making technology and fluorescent staining technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The "gold standard" fungal culture method:

[0068] 1. Turn on the ultraviolet light of the ultra-clean workbench for disinfection and sterilization half an hour in advance, and clean and disinfect the operator's clothes and hands.

[0069] 2. Take out the prepared culture medium and number it correspondingly to the gynecological sample.

[0070] 3. According to the corresponding number, open the plate and drop a drop of sterilized physiological saline on it, dip the sampling swab in physiological saline and spread it evenly on the plate, cover the petri dish, and put the sampling swab back to its original place. tube.

[0071] 4. Invert the petri dish and incubate in a biochemical incubator at 33°C for 24-72 hours, during which time observe the growth of the colony in the petri dish.

Embodiment 2

[0073] Ordinary fungal fluorescent staining method:

[0074] 1. Rotate the sample evenly to the middle of the slide,

[0075] 2. Add fungal fluorescent staining solution dropwise, cover with a cover glass, and examine under the microscope.

Embodiment 3

[0077] Fungal fluorescent staining method based on liquid-based film production technology

[0078] 1. Collect the sample and put the sample into the sample kit containing the cell preservation solution;

[0079] 2. Place the sample kit on the shaker and shake to mix;

[0080] 3. Install the sample filter and slide glass on the liquid-based fungal film maker;

[0081] 4. Add all the sample liquid in the sample kit to the installed sample filter;

[0082] 5. Start the liquid-based fungal film maker for automatic film production;

[0083] 6. Take out the slide after the production is completed, add 50ul fluorescent staining solution dropwise on the slide, cover it with a cover glass and let it stand for 30s;

[0084] 7. Perform microscopic examination under a fluorescent microscope.

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Abstract

The invention belongs to the field of microorganism detection, and particularly relates to a fungus detection method based on liquid-based slide making technology and fluorescent staining technology.A fluorescent staining solution is prepared from the following raw materials: 0.001% of fluorescent brightener 28, 10% of potassium hydroxide, 4% of dimethyl sulfoxide, 3.6% of sodium chloride, 10% ofglycerol, 0.01% of Evans blue, and the balance of purified water. Most of samples can be enriched together and made into slides through the liquid-based making technology, so that the positive detection rate is greatly improved. According to the fungus detection method based on liquid-based slide making technology and fluorescent staining technology, the fluorescent staining solution can make thestructures of mycelia and spores clearly visible; fungi are easy to identify, the sensitivity is high; the accuracy and the efficiency of the staining result interpretation are improved; and the detection rate is high, and the application range is wide.

Description

technical field [0001] The invention provides a method for detecting fungi based on liquid-based film-making technology and fluorescent dyeing technology, which belongs to the field of microorganism detection. Background technique [0002] Fungi belong to eukaryotes, without plastids, and their nutritional mode is absorption without phagocytosis. The cell wall contains chitin and β-glucan. It can cause human and animal diseases, called fungal diseases, and can also cause plant diseases, human allergic diseases and mycotoxin poisoning. Fungal invasion of the human body is often secondary to other diseases, such as chronic wasting diseases such as malignant tumors and diabetes; large doses of X-ray irradiation; immunosuppressants, etc. lead to low immune function and resistance; The reaction is mostly suppressed, and it is also easy to be infected with fungi; in addition, organ transplantation, catheter intubation, radiotherapy and other applications cause local tissue damag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/569
CPCG01N21/6428G01N33/56961G01N2021/6439
Inventor 余尧毅熊文芳吕正和陈永洪杜厚明杨海敏谢赦飞柯子璇
Owner 江西业力医疗器械有限公司
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