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POCT fluorescence quantitative detection method and detection kit for group A streptococcus

A technology of streptococci and fluorescent microspheres, applied in biological testing, fluorescence/phosphorescence, measuring devices, etc., can solve the problems of false negative sensitivity, inability to diagnose in time, complex operation, etc., achieve high specificity, reduce waste of medical resources, Simple operation effect

Inactive Publication Date: 2018-08-24
ZHUHAI MEIHUA MEDICAL TECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the domestic identification of GAS mainly relies on the bacterial culture method, which takes about 2 to 4 days, cannot be diagnosed in time, and has defects such as false negatives (that is, insufficient sensitivity) and complicated operations.

Method used

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  • POCT fluorescence quantitative detection method and detection kit for group A streptococcus
  • POCT fluorescence quantitative detection method and detection kit for group A streptococcus
  • POCT fluorescence quantitative detection method and detection kit for group A streptococcus

Examples

Experimental program
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Effect test

Embodiment 1

[0045] This embodiment 1 provides a POCT fluorescence quantitative detection kit for group A streptococci, such as figure 1 As shown, the POCT fluorescence quantitative detection kit includes a box body 1 and an immunochromatographic test strip disposed in the box body 1 .

[0046]Specifically, the immunochromatographic test strip includes a PVC sticky bottom plate 2 and a sample pad 3 fixed on the PVC sticky bottom plate 2, a bonding pad 4, an NC film 5 and a water-absorbing pad 6, and the bonding pad 4 is fitted and fixed on the PVC sticky bottom plate 2. Below the sample pad 3 , one end of the NC membrane 5 is attached and fixed below the bonding pad 4 , and the other end is connected to the water-absorbing pad 6 .

[0047] Specifically, a sampling port 7 is opened above the sample pad 3 on the surface of the box body 1 .

[0048] Specifically, fluorescent microsphere-labeled antibodies are sprayed on the binding pad 4, and the fluorescent microsphere-labeled antibodies ar...

Embodiment 2

[0054] Present embodiment 2 provides a kind of preparation method of the immunochromatographic test paper strip of detection group A streptococcus, comprises the steps:

[0055] (1) Coat group A streptococcus antibody and goat anti-mouse secondary antibody on the NC membrane with Bio-Dot XYZ3000 film spotting instrument, group A streptococcus antibody as the detection line, goat anti-mouse secondary antibody as the quality control line, 36 Dry overnight in a drying oven at -38°C; the group A streptococcal antibody is a mouse monoclonal antibody or a rabbit polyclonal antibody; specifically, the drying temperature of the NC film can be selected to be 37°C, which is approximately human body temperature;

[0056] (2) Spray the fluorescent microsphere-labeled antibody on the binding pad, and dry it in vacuum at 30°C for 2 hours;

[0057] (3) After the NC membrane, binding pad, sample pad, absorbent paper and PVC sticky bottom plate are assembled, cut into test strips, add a desicc...

Embodiment 3

[0059] This embodiment 3 provides a method for preparing fluorescent microsphere-labeled antibodies, comprising the following steps:

[0060] (1) Replace the original preservation solution of the microspheres with the activation solution: centrifuge, remove the supernatant, and ultrasonically resuspend the activation solution;

[0061] (2) Add EDC and Sulfo-NHS for activation: Weigh a certain amount of EDC and Sulfo-NHS, dissolve them, add them to the microsphere solution and rotate for 15-20 minutes;

[0062] (3) Remove unreacted or residual reactants: perform centrifugation, supernatant removal, and ultrapure water ultrasonic resuspension in sequence, repeating 2 to 3 times;

[0063] (4) Ultrasonically resuspend the microspheres in the reaction solution, then add a certain amount of group A streptococcus antibody to be labeled, and react in the dark for 1 to 4 hours;

[0064] (5) Removal of unconjugated group A streptococcal antibodies: perform centrifugation, supernatant r...

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Abstract

The invention discloses a POCT fluorescence quantitative detection method and a detection kit for group A streptococcus, and the POCT fluorescence quantitative detection kit comprises a box body and an immunochromatographic test strip disposed in the box body, the immunochromatographic test strip is coated with a group A Streptococcal antibody which is a mouse monoclonal antibody or a rabbit polyclonal antibody. At the time of detection, after a sample is added to the detection kit, a group A streptococcus antigen is sequentially combined with a fluorescent microsphere-labeled antibody and a detection antibody by the chromatography effect of the immunochromatographic test strip, finally a fluorescence signal is detected by a fluorescence detector, and because the fluorescence signal is positively correlated with the content of the group A Streptococcus antigen, the group A Streptococcus antigen in a throat swab sample can be quantitatively detected. The detection method is simple and rapid, and has high sensitivity and specificity, so that patients with GAS infection can get timely diagnosis and treatment, and the detection method can also provide a basis for rational use of antibiotics in the patients with GAS infection in clinical treatment.

Description

technical field [0001] The application relates to the field of antibodies, in particular to a POCT fluorescence quantitative detection method and detection kit for Group A Streptococcus. Background technique [0002] The full name of group A streptococcus is group A beta (β) hemolytic streptococcus, English Group A Streptococcus (abbreviated as Strep A or GAS), and Chinese is also called Streptococcus pyogenes, which is the most pathogenic bacteria in streptococci. Group A streptococci can produce a variety of enzymes and exotoxins, and are the most common pathogens that cause bacterial upper respiratory tract infections in children aged 3-15, and are extremely common in clinical work. Early diagnosis and appropriate antibiotic treatment within 48 hours after the onset of GAS infection are very effective; if not promptly and effectively treated, it may lead to acute necrotizing fascitis (ANF) and streptococcal toxic shock syndrome (streptococcal toxin shock syndrome, STSS) ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/533G01N33/577G01N33/558G01N33/68G01N21/64
CPCG01N21/64G01N21/6486G01N33/533G01N33/558G01N33/56944G01N33/577G01N33/6854G01N33/6893G01N2333/315
Inventor 乌月恒张志伟
Owner ZHUHAI MEIHUA MEDICAL TECH LTD