A kind of fungal fluorescent staining solution and preparation method thereof

Abstract

The invention discloses a fungal fluorescent staining fluid, which is prepared from, by weight (%), 0.08-0.012% of 5-carboxy flouorescein, 0.045-0.055% of fungal primer ITS4, 8-13% of potassium hydroxide, 28-33% of glycerinum, 0.45-0.55% of diphosphatidyl glycerol, 0.07-0.13% of ascorbic acid, 53.363-63.427% of water. Through adding 5-carboxy flouorescein and fungal primer, the fungal primer ITS4forms a stem ring structure under a free state, thus the fluorescent recession can be effectively avoided; 5-carboxy flouorescein in the of potassium hydroxide solution can penetrate through a fungalcell wall and a cell film to generate specificity combination with alkali group of the ribonucleic acid gene interval zone of fungal ribosome; at the moment, the stem ring structure is opened and shows fluorescence through microscopic examination under a fluorescence microscope; potassium hydroxide can smelt fungal cuticle in the dyeing process, thus the fungal primer ITS 4 can rapidly enter the internal action of the fungal cell; besides, for the fluorescent recession inhibitor is added in the formula, the fluorescence primer can be kept for a long term after the specificity combination withthe alkali group of the ribonucleic acid gene interval zone of fungal ribosome.

Description

technical field [0001] The invention relates to the technical field of biological testing of pathogenic bacteria in medical clinical testing, in particular to a fungal fluorescent staining solution for fungal cell staining and a preparation method thereof. Background technique [0002] In the current medical clinical examination, the traditional potassium hydroxide method or the immunological test method developed in recent years are mostly used in the detection of fungal infection. in: [0003] Although the potassium hydroxide method is relatively simple to operate, there are many shortcomings. One is that it is not specific and is easily affected by other impurity substrates, resulting in unclear observation and prone to false negatives; It is necessary to find fungal mycelium or spore structure to get the test conclusion, so it takes a lot of test time and the work efficiency is low; thirdly, the test range of this method is limited, and it does not work at all for Pityr...

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Problems solved by technology

[0003] Although the potassium hydroxide method is relatively simple to operate, there are many shortcomings. One is that it is not specific and is easily affected by other impurity substrates, resulting in unclear observation and prone to false negatives; It is necessary to find the fungal hyphae or spore structure to get the test conclusion, so it takes a lot of test time and the work efficiency is low; the third is that the test range of this method is limited, and it does not work at all for Pityrosporum and Malassezia

[0004] Although the immunological test method has good specificity, it can only be tested for antibodies against deep fungal infections. Its disadvantages are: First, it cannot be detected in time for systemic infections who do not have antibodies due to reduced immune system function. ; Second, for patients who have developed antibodies for a long time during the recovery from infection, there is a certain false positive rate in this group of people

[0005] With the continuous improvement of medical inspection level, fungal fluorescent staining clinical inspection technology has been gradually promoted in recent years. At present, most of the fluorescent staining methods such as cell wall β-polysaccharide and chitin are used in the market. This method only uses fluorescent dyes and cell wall β-polysaccharides, Chitin binds with affinity and has no specificity; with the gradual promotion of clinical practice, it is easily interfered by impurities in the dyeing substrate, which affects the judgment of observation results; in addition, the fluorescence decays quickly, and samples cannot be preserved; and applications causing great trouble

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