A rapid staining reagent for Leptospira and its preparation method and application

A leptospira and rapid staining technology, which is applied in biochemical equipment and methods, microbiological measurement/inspection, and resistance to vector-borne diseases, etc., can solve the problems of cumbersome dyeing process, long dyeing time, and difficult observation, and achieve operational Less skill required, shorter staining time, more visible results

Active Publication Date: 2020-07-31
XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the Fontana silver-plating staining inspection method is usually used. This method requires expensive staining reagents and needs to be prepared and used immediately. The prepared reagents are unstable and cannot be stored for a long time. The staining process is cumbersome, and the silver-plated staining time is difficult to control. No staining, long staining time, silver particles often precipitate in the specimen, more impurities, difficult to observe
Due to the oxidation of silver ions, the specimen slices dyed by silver-plating staining method are unstable and cannot be stored for a long time

Method used

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  • A rapid staining reagent for Leptospira and its preparation method and application
  • A rapid staining reagent for Leptospira and its preparation method and application
  • A rapid staining reagent for Leptospira and its preparation method and application

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Effect test

Embodiment 1

[0020] A rapid staining reagent for Leptospira, made from the following raw materials: 36g (30ml) of lactic acid, 3.2g of aluminum chloride, 2.2g of sucrose, 1.28g of malachite green, 17.38g of absolute ethanol (22ml), 50g of distilled water ( 50ml), 0.2g of 1mol / L NaOH aqueous solution.

[0021] The preparation method of described Leptospira rapid staining reagent comprises the following steps:

[0022] (1) Put the malachite green dye in a mortar, grind it thoroughly, add absolute ethanol to fully dissolve it, and filter it with two layers of filter paper to obtain the filtrate;

[0023] (2) Dissolve aluminum chloride in distilled water, then add sucrose and lactic acid and mix well to obtain solution A;

[0024] (3) Mix the filtrate obtained in step (1) with solution A obtained in step (2), mix well, add 1mol / L NaOH aqueous solution, mix well, store in a brown jar, and store at room temperature for later use.

Embodiment 2

[0026] A rapid staining reagent for Leptospira, made from the following raw materials: 31.2g (26ml) of lactic acid, 3.65g of aluminum chloride, 4.3g of sucrose, 1.59g of malachite green, 14.22g (18ml) of absolute ethanol, and 56g of distilled water (56ml), 0.5g of 1mol / L NaOH aqueous solution.

[0027] The preparation method of described Leptospira rapid staining reagent comprises the following steps:

[0028] (1) Put the malachite green dye in a mortar, grind it thoroughly, add absolute ethanol to fully dissolve it, and filter it with two layers of filter paper to obtain the filtrate;

[0029] (2) Dissolve aluminum chloride in distilled water, then add sucrose and lactic acid and mix well to obtain solution A;

[0030] (3) Mix the filtrate obtained in step (1) with solution A obtained in step (2), mix well, add 1mol / L NaOH aqueous solution, mix well, store in a brown jar, and store at room temperature for later use.

Embodiment 3

[0032] A rapid staining reagent for Leptospira, made from the following raw materials: 24g (20ml) of lactic acid, 3.2g of aluminum chloride, 5.3g of sucrose, 2.65g of malachite green, 11g of absolute ethanol (13.9ml), 48g of distilled water ( 48ml), 0.1g of 1.2mol / L NaOH aqueous solution.

[0033] The preparation method of described Leptospira rapid staining reagent comprises the following steps:

[0034] (1) Put the malachite green dye in a mortar, grind it thoroughly, add absolute ethanol to fully dissolve it, and filter it with two layers of filter paper to obtain the filtrate;

[0035] (2) Dissolve aluminum chloride in distilled water, then add sucrose and lactic acid and mix well;

[0036] (3) Mix the filtrate obtained in step (1) with the solution obtained in step (2), mix well, add 1.2mol / L NaOH aqueous solution, mix well, store in a brown jar, and store at room temperature for later use.

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Abstract

The invention belongs to the technical field of microbiological detection, and in particular relates to a spirochete dyeing reagent and its application. The invention provides a new dyeing reagent and a method of use thereof for Leptospira dyeing. The preparation of the reagent is simple, and the cost of materials used in the preparation of the reagent is low. Easy to obtain, the prepared reagent does not require special storage conditions, and can be stored at room temperature for a long time without failure. The staining method is simple to operate and does not require high skills for the experimenter; and the staining time only takes 40-60 seconds, which greatly shortens the staining time. time, the staining effect is good, and the Leptospira dyed by this method is green, which is easier to observe under the microscope and has no impurities. It is suitable for promotion and application in staining of Leptospira.

Description

technical field [0001] The invention belongs to the technical field of microbiological detection, and in particular relates to a spirochete staining reagent and an application thereof. Background technique [0002] Leptospira belongs to the genus Leptospira in the Leptospira family of the order Spirochetes. The thallus is slender, 6-12 microns long and 0.1-0.2 microns wide. One or both ends of the thallus are bent so that the thallus is in the shape of a question mark. The disease caused by pathogenic Leptospira is leptospirosis, referred to as leptospirosis, which is a typical zoonotic disease with a global distribution. In my country, except Xinjiang, Tibet, Qinghai, and Ningxia Leptospirosis is prevalent in other areas except Gansu and Gansu (Edited by Li Fan and Xu Zhikai, "Medical Microbiology, Eighth Edition, P198~P208), so this disease is a key point of prevention and control in my country one of infectious diseases. Due to the difference in the serotype, virulence an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04
CPCC12Q1/04Y02A50/30
Inventor 杨帆李敏马玉龙邓保国魏纪东
Owner XINXIANG MEDICAL UNIV
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