Kit for quantitatively detecting trichomonas vaginalis by chemiluminiscence method and detecting method thereof

A Trichomonas vaginalis, chemiluminescence technology, applied in chemiluminescence/bioluminescence, color/spectral property measurement, analysis by chemical reaction of materials, etc., can solve the requirements of high experimental conditions, pollution, sensitivity and The problem of high specificity can achieve the effect of high detection sensitivity and accuracy, reducing non-specific binding and improving reaction speed.

Inactive Publication Date: 2019-11-22
江苏美克医学技术有限公司
View PDF5 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Smear staining method is not sensitive enough and interferes with particles similar in size to Trichomonas
The culture method is considered as the "gold standard" for diagnosing trichomonas with high sensitivity, but it takes a long time to culture and has few clinical applications
[0005] Molecular biology methods, mainly using nucleic acid molecular hybridization technology or PCR technology, have high sensitivity and specificity, but they have high requirements for experimental conditions, there may be pollution problems, and the cost is high

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Kit for quantitatively detecting trichomonas vaginalis by chemiluminiscence method and detecting method thereof
  • Kit for quantitatively detecting trichomonas vaginalis by chemiluminiscence method and detecting method thereof
  • Kit for quantitatively detecting trichomonas vaginalis by chemiluminiscence method and detecting method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Preparation of neutravidin-coupled magnetic microspheres

[0063] Take 10-100mM activation buffer and wash 10 mg of magnetic microspheres twice; the activation buffer is fatty acid methyl ester sulfonate (MES) with a pH of 5.0-7.0;

[0064] Add 1-10mg EDC and 1-10mg NHS solution to activate the magnetic microspheres, mix well, and react on a rotary mixer at 37°C for 15min-1h at a speed of 30-60hr, remove the supernatant by magnetic adsorption; then use 50mM pH 7.5- Wash twice with 8.5 HEPES to obtain activated magnetic microspheres;

[0065] Add 0.5-1 mg of neutravidin and react on a rotary mixer for 2-3 hours;

[0066] Add 0.05-0.2mol / L glycine and 0.05-0.2mol / L ethanolamine solution to block overnight;

[0067] The next day, remove the supernatant by magnetic adsorption, wash three times with 10-50mM PBST, add microsphere preservation solution to preserve the magnetic microspheres to obtain the neutravidin-coupled magnetic microspheres, and store them at 2-8°C for l...

Embodiment 2

[0069] Preparation of biotinylated antibodies

[0070] Take sodium bicarbonate buffer solution with a pH of 8.0-9.0 and fully dialyze 0.5mg of antibody at a concentration of 1-2mg / ml, add NHSB, wherein the molar ratio of NHSB:antibody is 20-100:1, and react at 37°C for 2-4h, 1 mol / L ammonium chloride solution was added to block for 10-30 min; the biotinylated antibody was collected by dialyzing overnight with PBS buffer at pH 7.4.

Embodiment 3

[0072] Preparation of acridinium ester-labeled antibodies

[0073] After fully dialyzing the Trichomonas vaginalis antibody with carbonic acid buffer (pH 9.0-10.0), add acridinium ester (the molar ratio of acridinium ester to antibody is 10:1, 15:1, 20:1), and react at room temperature for 2 hours , blocked with lysine solution, dialyzed overnight in 10mM PBS (pH7.4) buffer solution, collected the reaction product, and obtained acridinium ester-labeled antibody. The labeling effects of different acridinium ester labeling amounts are shown in Table 1.

[0074] Table 1 Marking effect of different acridinium ester labeling amounts

[0075]

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a kit for quantitatively detecting trichomonas vaginalis by a chemiluminiscence method. In use, a trichomonas vaginalis antigen in a sample reacts with a reagent in the kit to generate a neutral avidin microsphere-capture antibody-antigen-detection antibody-acridinium ester compound; washing is carried out to remove substances which do not participate in the reaction; and when acridinium ester is in an alkaline H2O2 solution, molecules are attacked by hydrogen peroxide ions, unstable dioxane is generated, the dioxane is decomposed into CO2 and N-methyl acridone in an electron excited state, photons are emitted when acridinium ester returns to a ground state, and a light-emitting signal measuring instrument is used for measuring the light quantum yield for measurement. Meanwhile, a quantitative standard curve is fitted through a calibrator, and the concentration of trichomonas vaginalis is calculated by substituting the light signal measurement result of the sample into the quantitative standard curve. The kit provided by the invention is used for quantitative detection of trichomonas vaginalis, the sensitivity is high, the specificity is good, and the concentration of trichomonas vaginalis in the sample can be accurately detected.

Description

technical field [0001] The invention belongs to the technical field of in vitro diagnosis, and in particular relates to a kit for quantitatively detecting Trichomonas vaginalis by chemiluminescence and a detection method thereof. Background technique [0002] Trichomonias is a flagellated protozoan organism that parasitizes the human urogenital tract, can cause vaginitis and urethritis, and is one of the risk factors for HIV infection and cervical cancer. Women infected with Trichomonas vaginalis in the second trimester are prone to premature birth and low birth weight babies. There are literatures that trichomonas infection is one of the important causes of male nongonococcal urethritis. With an annual incidence of 180 million Trichomonas vaginalis infections worldwide, trichomoniasis is considered the most prevalent sexually transmitted disease. [0003] At present, trichomoniasis detection methods mainly include etiological diagnosis, immunological methods, molecular bi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N21/31G01N33/577G01N33/569G01N33/543
CPCG01N21/31G01N21/76G01N33/54306G01N33/54326G01N33/56905G01N33/577
Inventor 邓玲玲
Owner 江苏美克医学技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap