Pathogenic bacterium liquid-based thin-layer smear detection kit and detection method thereof

A detection kit and detection method technology, which are applied in the preparation of test samples, biochemical equipment and methods, and microbial determination/inspection, etc., can solve the problem of quickly and sensitively providing pathogenic infection detection results for samples that cannot be pathogenic bacteria.

Inactive Publication Date: 2011-02-16
贺坚慧
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to solve the current clinical problem of not being able to quickly and sensitively provide pathogen infection detection results for samples containing pathogenic bacteria, the present invention improves on the above-menti

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Liquid-based thin-layer smear detection of mycobacterium tuberculosis in sputum (or cerebrospinal fluid or joint fluid) in embodiment 1

[0042] The sputum (or cerebrospinal fluid or joint fluid) samples of 200 cases of tuberculosis patients were taken to conduct a comparative experiment between liquid-based thin-layer smear method and direct smear method. The liquid-based thin-layer smear test kit for Mycobacterium tuberculosis is a pretreatment solution (100ml mixed solution prepared from 30ml of ammonia water with a density of 0.88g / ml, 4g of sodium chloride and the balance of water), with a concentration of 20mg The liquefaction agent of / ml mercaptoethanol, the concentration is the diluent of 0.9% sodium chloride solution, including the acid-fast staining solution and consumables of three reagents, carbofusin solution, 5% hydrochloric acid alcohol solution and methylene blue solution: (a) centrifugal precipitation Tablet assembly (Changsha Intech Instrument Co., Lt...

Embodiment 2

[0044] Example 2 Pseudomonas aeruginosa liquid-based thin-layer smear detection in pus and wound infection tissue samples

[0045] The liquid-based thin-layer smear detection kit for Pseudomonas aeruginosa is a pretreatment solution (100ml mixed solution prepared by 25ml of ammonia with a density of 0.88g / ml, 1g of sodium chloride and the balance being water), with a concentration of The liquefaction agent of 5.0mg / ml dithioerythritol, the concentration is the diluent of 1% sodium chloride solution, the concentration is the antibody solution of 0.5 μ g / ml chicken anti-Pseudomonas aeruginosa antibody (IgY)-HRP, chromogen (The chromogen comprises a concentration of 0.40mg / ml DAB substrate solution, a concentration of 0.35% by weight hydrogen peroxide and a concentration of 10mg / ml cobalt chloride solution) and consumables, consumables and equipment used are the same as in Example 1.

[0046]There are three situations for pus and wound infected tissue samples: 1. For open infecti...

Embodiment 3

[0047] Embodiment 3 liquid-based thin-layer smear detection of Staphylococcus aureus and / or Streptococcus pneumoniae and / or Candida albicans in sputum (or bronchial lavage) samples

[0048] The detection kit used for the joint detection of Staphylococcus aureus, Streptococcus pneumoniae and Candida albicans is 100ml prepared by the pretreatment liquid (by 37ml of ammonia water with a density of 0.88g / ml, 5g of sodium chloride and the balance being water). mixed solution), the concentration is the liquefaction agent of 22mg / ml β-mercaptoethanol, the diluent of 1.5% sodium chloride solution, the Gram staining solution including crystal violet, iodine solution, decolorant and counterstaining agent , a concentration of 0.4 μg / ml rabbit IgG-HRP antibody solution and a chromogenic agent (the chromogenic agent includes a concentration of 0.75 mg / ml DAB substrate solution, a concentration of 0.67 weight ‰ hydrogen peroxide and a concentration of 8 mg / ml cobalt chloride solution ) and ...

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Abstract

The invention relates to a pathogenic bacterium liquid-based thin-layer smear detection kit and a detection method thereof. The detection kit comprises a preconditioning liquid, a liquefier, a diluent, a differential staining liquid and consumable items. The detection method of the detection kit comprises the following steps: adding the strongly alkaline preconditioning liquid and the liquefier into a detection sample to carry out a homogeneous treatment on the sample, adding the diluter, adopting a liquid-based centrifugal settling thin-layer preparation technology to obtain multiple uniformly-distributed pathogenic bacterium liquid-based thin-layer smears, carrying out differential staining, observing and calculating according the staining reaction, the morphology and the arrangement mode of the pathogenic bacteria, thus working out a detection result of multiple pathogenic bacteria in the sample. The invention establishes a fast and sensitive detection method based on the pathogenic bacterium liquid-based thin-layer smears, the whole process can be completed within one hour, and the detection method is simple, thereby achieving high clinical popularization values.

Description

technical field [0001] The invention relates to a liquid-based thin-layer smear detection kit for pathogenic bacteria and a detection method thereof. The detection method of this kit is mainly to homogenize the detection sample with a pretreatment solution and a liquefaction agent, then dilute it with a diluent, and then make a centrifugal sedimentation sheet to obtain multiple liquid-based thin-layer smears of pathogenic bacteria, and then Differential staining can be used to quickly detect a variety of pathogenic bacteria in the sample according to the staining reaction, morphology and arrangement of the pathogenic bacteria. Background technique [0002] Respiratory infection is a relatively common disease. Among the various pathogens of lung infection, bacteria account for the first place (about 80% for adults and about 60% for children), followed by viruses, mycoplasma, etc. Medical Science Press, PP 974-981). Streptococcus pneumoniae, which is more important in upper...

Claims

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Application Information

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IPC IPC(8): G01N1/28C12Q1/04G01N1/30
CPCY02A50/30
Inventor 贺坚慧潘曙明程振球吴增斌刘佳江静
Owner 贺坚慧
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