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Combined detection kit for detecting and identifying candida and trichomonias

A kit and Candida technology, applied in the field of joint inspection kits for detecting and identifying Candida and Trichomonas, can solve the problems of high impact, no high-quality products, and large impact of paraffin oil in the field of vision, so as to reduce subjective factors. Influence, objective and reliable test results, convenient and quick use method

Active Publication Date: 2014-08-13
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is greatly affected by experience, and is greatly affected by the field of view, interfering particles, especially the paraffin oil added for lubricating the specimen.
2. Culture method: This method is very sensitive, but it takes a long time to produce results, and the timeliness is poor, which may delay the disease.
3. Latex agglutination slide test: use polyclonal antibody to react with yeast mannan of some candida, but no high-quality products that can be used clinically
The hanging drop method is the easiest way to check Trichomonas vaginalis, but there is a problem of low sensitivity, because this method detects Trichomonas by observing the motility of Trichomonas, and the motility of Trichomonas is greatly affected by temperature. Significantly decreased mobility
2. Smear staining method: smear the secretion on a glass slide, and after natural drying, it can be stained with different dye solutions, such as Gram staining, etc., but this method is interfered by particles similar in size to Trichomonas and affected by the field of view
At present, the N-acetyl-hexosaminidase method can simultaneously detect and distinguish Trichomonas and Candida, but the sensitivity and specificity of this method are low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] 1. Preparation of kits

[0028] 1. It is called glycyl-arginine-4-methoxy-β-naphthylamine hydrochloride (Gly-Arg-4-methoxy-β-naphthylamine HCl, produced by Sigma-Aldrich Company) (glycine can also be used acyl-arginine-β-naphthylamine), dissolved in 400mM phosphate buffer (pH4.0-8.0) to make the concentration reach 2-5mg / ml; Aminobenzaldehyde) 0.2g dissolved in 1000ml 0.3mol / L H 2 SO 4solution; the volume ratio of the above two solutions is 1:1, mix evenly, add 10ul / piece to a 5mm diameter circular paper, and dry overnight to obtain coagulase detection test paper.

[0029] 2. It is called 6-chloro-3 indole-a-D-glucopyranoside (manufactured by Cabosens Chemical Technology (Suzhou) Co., Ltd.) (5-Br-6-chloro-3 indole-a-D-pyridine can also be used glucopyranoside, 5-Br-4-chloro-3-indole-a-D-glucopyranoside), dissolved in an appropriate amount of methanol, and then dissolved in 400mM phosphate buffer (pH5.0) to a concentration of 0.5 -2mg / ml, mix well. Add 10ul / sheet to...

Embodiment 2

[0039] 1. Kit preparation

[0040] 1. Z-glycyl-arginine-p-nitroanilide (Z-Gly-L-Arg-4-nitroanilide, produced by Shanghai Hanhong Chemical Co., Ltd.), with 400mM phosphate buffer (pH4.0) Dissolve to make the concentration reach 5mg / ml; Add 10ul / sheet to a 5mm diameter circular paper, dry overnight to obtain coagulase test paper.

[0041] 2. It is called 6-Br-2-naphthol-a-D-glucopyranoside (manufactured by Cabosens Chemical Technology (Suzhou) Co., Ltd.) (6-Br-1-naphthol-a-D-glucopyranose can also be used Glycosides), dissolved in appropriate amount of ethanol, then dissolved in 400mM phosphate buffer (pH5.5) to make the concentration reach 2mg / ml, and mix well. Add 10ul / sheet to a circular paper sheet with a diameter of 5mm, and dry overnight to obtain α-glucosidase detection test paper.

[0042] 3. Weigh the absolute ethanol solvent for bromocresol green, mix it evenly to make the concentration reach 2mg / ml, add 15μl / piece to a paper piece with a side length of 5mm, and dry ...

Embodiment 3

[0051] One, kit preparation of the present invention

[0052] 1. Glycyl-arginine-7-amino-4-methylcoumarin (Gly-L-Arg-7-amino-4-methylcoumarin, produced by Shanghai Gil Biochemical Co., Ltd.), with 100-400mM Tris -Dissolve in HCl buffer (pH8.0) to make the concentration 0.2-1mg / ml; add 10ul / sheet onto a 5mm diameter disc, and dry overnight to obtain coagulase test paper.

[0053] 2. It is called 4-methylumbelliferone-α-D-glucopyranoside (manufactured by Cabosens Chemical Technology (Suzhou) Co., Ltd.) (you can also use vanillin-α-D-glucopyranoside, 4 -methylcoumarin-α-D-glucopyranoside, 4-methylcoumarin-2-deoxy-2-sulfate amino-α-D-glucopyranoside sodium), with an appropriate amount of dimethyl After the sulfone is dissolved, dissolve it in 100mM-400mM acetate buffer (pH5.1) to make the concentration reach 1mg / ml, and mix well. Add 10ul / sheet to a circular paper sheet with a diameter of 5mm, and dry overnight to obtain a-glucosidase detection test paper.

[0054] 3. Weigh the...

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Abstract

The invention discloses a joint detection kit for detecting and distinguishing Trichomonas and Candida, which comprises coagulation enzyme activity detection reagent, α-glucosidase activity detection reagent and pH detection reagent; the enzyme substrate used in the coagulation enzyme activity detection reagent The substance is a Gly-Arg dipeptide substrate containing a chromogen or a fluorescent group; the enzyme substrate used in the α-glucosidase activity detection reagent is an α-D-glucoside substrate containing a chromogen or a fluorescent group . In order to improve the color developing effect, it may also include a color developing solution. The invention has the advantages of relatively high sensitivity and specificity, significantly improved performance of reagents compared with the existing similar methods, and can meet the needs of clinical detection.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a joint detection kit for detection and identification of candida and trichomonas. Background technique [0002] Candida (Candida) is the most common opportunistic pathogen in fungi, also known as Candida, is one of the main pathogens that cause human urogenital diseases, mainly including Candida albicans, Candida glabrata, Candida tropicalis and Gram Candida bruisei, etc. Under normal circumstances, Candida parasitizing in the human body is of yeast cell type, and generally does not cause disease. Under the influence of certain physiological and pathological factors of the body, when the internal environment of the vagina / penis changes and the body’s resistance / immunity decreases, Candida will multiply and develop into hyphae, invade tissues, and cause candidiasis, so Candida It is an opportunistic pathogen. These factors usually include pregnancy, diabetes, oral...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/34C12Q1/25C12Q1/04G01N27/00
Inventor 杨红云刘志磊王敏王则宇杨静静石乐秦磊张硕张利娟杨晓华耿晓东付光宇王胜启孙若楠吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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