Improved mycobacterium tuberculosis rapid anti-acid fluorescence staining reagent

A fluorescent staining technology for Mycobacterium tuberculosis, applied in the field of biomedical diagnosis, can solve the problems of delayed treatment, increased risk, and long detection cycle, and achieve the effects of good stability, shortened staining time, and convenient clinical use

Inactive Publication Date: 2018-12-04
江苏诺鬲生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the culture method has high sensitivity, the detection cycle is too long, whic...

Method used

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  • Improved mycobacterium tuberculosis rapid anti-acid fluorescence staining reagent
  • Improved mycobacterium tuberculosis rapid anti-acid fluorescence staining reagent
  • Improved mycobacterium tuberculosis rapid anti-acid fluorescence staining reagent

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1 Reagent 1: the preparation of the patent reagent of the present invention

[0027] Solution A: Dissolve auramine O powder in a solution with a concentration of 0.9% (W / V) sodium chloride, fully dissolve, so that the concentration of auramine O is 0.1% (W / V), and then add dimethyl methylene Sulfone and glycerol were used so that the concentration of dimethyl sulfoxide was 15% (v / v) and the concentration of glycerol was 20% (v / v).

[0028] Solution B: Prepare the acid alcohol solution first, so that the concentration of hydrochloric acid is 0.3% (V / V), and the concentration of ethanol is 75% (V / V). Then add methyl green to the acid-alcohol solution so that the concentration is 0.5% (W / V).

[0029] Reagent 2: Configuration of traditional auramine O:

[0030] Solution A: Dissolve 0.1g of auramine O powder in 10ml of 95% ethanol, add diluted 5% carbolic acid to 100ml;

[0031] Solution B: Prepare an acid alcohol solution so that the concentration of hydrochlo...

Embodiment 2

[0033] Example 2 Staining of sputum specimens

[0034] Select sputum samples positive for Mycobacterium tuberculosis, spread them on two glass slides, and stain with reagent 1 and reagent 2, respectively. Reagent 1 operation steps: After staining with solution A for one minute, rinse with water, stain with solution B for one minute, mount the slides and observe under a microscope after washing. Reagent 2 operation steps: stain with reagent A for 15 minutes, rinse with water, stain with solution B for 2 minutes, counterstain with solution C for 1 minute, cover the slides, absorb the excess dye with paper, and then stain in the fluorescence. Observe under a microscope. When observed under the blue light band of 410 nm–440 nm, Mycobacterium tuberculosis shows green fluorescence. The results show that both reagents can make tubercle bacillus label fluorescent, and the staining time of the reagent of the present invention is shorter.

Embodiment 3

[0035] Example 3 Staining of lung tissue paraffin sections

[0036] Select three paraffin-embedded section specimens, and stain with reagent 1 and reagent 2 after dewaxing. Reagent 1 operation steps: after staining with solution A for one minute, rinse with water, stain with solution B for one minute, and mount the slide under microscope after washing. Next observe. Reagent 2 operation steps: stain with reagent A for 15 minutes, rinse with water, stain with solution B for 2 minutes, counterstain with solution C for 1 minute, cover the slides, absorb the excess dye with paper, and then stain in the fluorescence. Observe under a microscope. When observed under the blue light band of 410 nm–440 nm, Mycobacterium tuberculosis shows green fluorescence. The results show that both reagents can make tubercle bacillus label fluorescent, and the staining time of the reagent of the present invention is shorter.

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Abstract

The invention discloses an improved mycobacterium tuberculosis rapid anti-acid fluorescence staining reagent comprising two reagents: an A liquid, comprising pure water, NaCl or KCl, dimethyl sulfoxide, glycerol and auramine O; and a B liquid, comprising pure water, hydrochloric acid, ethanol, and methyl green or Evans blue. Through formula optimization, auramine O fluorescence staining method isimproved and optimized, and a double-reagent type mycobacterium tuberculosis rapid anti-acid fluorescence staining reagent is developed in the invention, by which staining is completed just through two steps, and staining time is reduced from 20 min, in conventional processes, to about 2 min. The staining reagent is more convenient and quick to apply clinically and greatly increases the detectionefficiency of the mycobacterium tuberculosis.

Description

technical field [0001] The invention belongs to the technical field of biomedical diagnosis, and in particular relates to an improved rapid acid-fast fluorescent staining reagent for mycobacterium tuberculosis. Background technique [0002] Pulmonary tuberculosis is a common infectious disease of the respiratory system, which spreads quickly. If it is not diagnosed and treated in time, it is easy to cause Mycobacterium tuberculosis to infect other organs or systems, such as bone tuberculosis, intestinal tuberculosis, and kidney tuberculosis. It not only brings great pain to patients, but also increases the difficulty of treatment, accelerates the progression of the disease, prolongs the treatment time, and increases the risk of organ failure. With the continuous in-depth development of my country's medical level, the gradual strengthening of the prevention and control of tuberculosis, and the gradual improvement of the public's medical and health requirements, under the comb...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N21/64
CPCG01N1/30G01N21/6486G01N2001/302
Inventor 何元林
Owner 江苏诺鬲生物科技有限公司
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