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A kind of cryptococcal capsule staining solution and its preparation and use method

A technique for capsular staining and cryptococcus, applied in biochemical equipment and methods, methods based on microorganisms, instruments, etc., can solve problems such as difficult operation, low positive rate, and inability to stain cryptococcus, achieving high test success rate, Increased positive rate and vivid coloring effect

Active Publication Date: 2021-09-24
马爽
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. Ink cannot color Cryptococcus, and the structure of Cryptococcus spores and capsules is not clear
[0006] 2. The test method lacks standards, and the capsule cannot be seen if it is not well grasped. Cryptococcus is easy to miss, which makes people feel difficult to operate and the positive rate is low.
[0007] 3. Indian ink is expensive

Method used

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  • A kind of cryptococcal capsule staining solution and its preparation and use method
  • A kind of cryptococcal capsule staining solution and its preparation and use method
  • A kind of cryptococcal capsule staining solution and its preparation and use method

Examples

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specific Embodiment 1

[0027] A kind of cryptococcal capsular staining solution provided by the present embodiment, its raw material comprises the following components:

[0028] 500 ml of methanol, 500 ml of phosphate buffered saline, 1 gram of Wright's powder, 0.3 gram of Giemsa powder, 10 gram of gum arabic and 10 gram of soot.

[0029] The phosphate buffer is a phosphate buffer with a pH value of 6.4 to 6.8; and the preparation method of the phosphate buffer is: dispersing 6.63 grams of anhydrous potassium dihydrogen phosphate and anhydrous phosphoric acid in every 1000 milliliters of pure water Disodium hydrogen 2.56 grams.

[0030] A kind of preparation method of cryptococcal capsule staining solution provided by the present embodiment comprises the following steps:

[0031] First, 1 gram of the Wright's dye powder and 0.3 gram of the Giemsa powder were dispersed in 500 milliliters of the methanol, then 500 milliliters of the phosphate buffer was added and mixed, and finally the methanol and t...

specific Embodiment 2

[0041] A kind of cryptococcal capsular staining solution provided by the present embodiment, its raw material comprises the following components:

[0042] 333 ml of methanol, 667 ml of phosphate buffered saline, 0.1 g of Crescendo, 0.03 g of Giemsa powder, 2 g of gum arabic, and 2 g of soot.

[0043] The preparation and use method of a cryptococcal capsule staining solution provided in this example can refer to specific example 1, except that the centrifuged sediment of the sample to be tested and the cryptococcal capsule staining solution are used according to the volume of 5 / 1 Than mixing, all the other steps are identical with specific embodiment 1 except this.

[0044] The observation results under the microscope after staining Cryptococcus in this example are similar to that of Example 1, but the clarity and contrast are not as good as those of Example 1.

specific Embodiment 3

[0046] A kind of cryptococcal capsular staining solution provided by the present embodiment, its raw material comprises the following components:

[0047] 667 ml of methanol, 333 ml of phosphate buffered saline, 10 g of Wright powder, 3 g of Giemsa powder, 20 g of gum arabic, and 20 g of soot.

[0048] The preparation and use method of a cryptococcal capsule staining solution provided in this example can refer to specific example 1, except that the centrifuged sediment of the sample to be tested and the cryptococcal capsule staining solution are used according to the volume of 1 / 5 Than mixing, all the other steps are identical with specific embodiment 1 except this.

[0049] The observation results under the microscope after staining Cryptococcus in this example are similar to those in Example 1, and the particles in some backgrounds become thicker and precipitate out, and the contrast is lower than that in Example 1.

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Abstract

The invention discloses a cryptococcal capsular staining liquid, the raw material of which comprises the following components: methanol, phosphate buffer, Wright's dye powder, Giemsa dye powder, gum arabic and soot, the methanol and the The mixing volume ratio of the phosphate buffer is 1 / (0.5~2), and the phosphate buffer is a phosphate buffer with a pH value of 6.4~6.8; every 1000 ml of the methanol and the phosphate buffer 0.1-10 grams of the Wright's dye powder, 0.03-3 grams of the Giemsa dye powder, 2-20 grams of the gum arabic, and 2-20 grams of the soot are dispersed in the mixture. The invention makes the spores of the cryptococcus brightly colored, the internal structure of the spores is clear, and is easier to identify; the standard dyeing method has a high test success rate; India ink is not needed; the dyeing liquid has stable properties and can be stored for a long time.

Description

technical field [0001] The invention relates to the technical field of cryptococcus detection, in particular to a cryptococcus capsule staining solution and a method for preparing and using the same. Background technique [0002] Cryptococcus is a fungus with a hypertrophic capsule, and its spore and capsule, capsule and peripheral edge are distinct, which is unique. Therefore, when people are suspected of being infected with Cryptococcus neoformans, observing the capsule of Cryptococcus becomes the first choice and rapid test, and the test results can be used for disease diagnosis, which is not the case at home and abroad. The method of observing the capsule is the so-called ink staining - "negative staining method", which has not changed. [0003] The characteristic of the negative staining method is: take cryptococcal suspension, cerebrospinal fluid or other body fluids, add Indian ink (black background), observe under a microscope, the cryptococcal capsule is not colore...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12R1/645
CPCC12Q1/04G01N2333/37
Inventor 马爽
Owner 马爽
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