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Influenza virus real-time detection kit and method

A technology for real-time detection of influenza virus, applied in biochemical equipment and methods, and microbial measurement/inspection, etc., can solve the problems of unsatisfactory timeliness, time-consuming, cumbersome reaction steps, etc., and achieve the reduction of automatic cracking degree and cost reduction , the effect of simplifying the operation steps

Inactive Publication Date: 2016-01-27
CELLEX BIOLOGICAL TECH SUZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to provide a one-step reaction step for influenza virus neuraminidase activity in order to solve the problems that the two-step reaction steps in the detection of influenza virus neuraminidase are relatively cumbersome, time-consuming, and unsatisfactory in timeliness. Reactive, real-time assay kit for rapid, real-time detection of influenza virus

Method used

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  • Influenza virus real-time detection kit and method
  • Influenza virus real-time detection kit and method

Examples

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Effect test

Embodiment 1

[0077] 1. The influenza virus real-time detection kit of the present invention, comprising a detection composition.

[0078] The detection composition formula is:

[0079]

[0080]

[0081] N-acetylneuraminic acid-firefly luciferin, the following formula 24μg / mL;

[0082]

[0083] Firefly luciferase 10 μg / mL.

[0084] Preparation method: mix the stock solution of each reagent in the above detection composition formula with deionized water, then adjust the pH value of the mixed solution to 7.0 with 1N NaOH or KOH solution, and then adjust the solution to 7.0 with deionized water to the final volume. Store overnight at 2-8°C.

[0085] Alternatively, divide the assay composition into aliquots of 0.5 mL (or other desired volumes) and fill into freeze-dried vials of appropriate capacity. Aliquots of the solution are then lyophilized and stored at an appropriate temperature (eg, not exceeding -20°C for long-term storage). When detecting influenza virus, one bottle of l...

Embodiment 2

[0092] 1. A real-time detection kit for influenza virus, including a detection composition, a capture reagent, a washing buffer, and a virus lysis buffer.

[0093] a. Detection composition

[0094] formula:

[0095]

[0096] N-acetylneuraminic acid-firefly luciferin, the following formula, 24 μg / mL;

[0097]

[0098] Firefly luciferase 10 μg / mL.

[0099] Preparation method: mix the stock solution of each reagent in the above detection composition formula with deionized water, then adjust the pH value of the mixed solution to 7.0 with 1N NaOH or KOH solution, and then adjust the solution to 7.0 with deionized water to the final volume. Store overnight at 2-8°C.

[0100] Alternatively, divide the assay composition into aliquots of 0.5 mL (or other desired volumes) and fill into freeze-dried vials of appropriate capacity. Aliquots of the solution are then lyophilized and stored at an appropriate temperature (eg, not exceeding -20°C for long-term storage).

[0101] b. ...

Embodiment 3

[0125] 1. A real-time detection kit for influenza virus, including a detection composition.

[0126] Detection composition formula:

[0127]

[0128] 4,7-methyl-N-acetylneuraminic acid-firefly luciferin, the following formula, 24μg / mL;

[0129]

[0130] Firefly luciferase 10 μg / mL.

[0131] Preparation method: mix the stock solution of each reagent in the above detection composition formula with deionized water, then adjust the pH value of the mixed solution to 7.2 with 1N NaOH or KOH solution, and then adjust the solution to 7.2 with deionized water. to the final volume. Store overnight at 2-8°C.

[0132] 2. Detection method

[0133] Mix 10 μL of the nasal rinse sample to be tested with 90 μL of the prepared detection composition, put the detection tube into the photometer and record the light signal (RLU) within 2 minutes. If there is a light signal, it means that there is influenza virus.

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Abstract

The invention discloses an influenza virus real-time detection kit and a method. The real-time detection kit comprises a detection composition, the pH value of the detection composition is 7.0-7.2, and the detection composition comprises N-acetylneuraminic acid-firefly luciferin conjugate or a salt thereof and firefly luciferase. The real-time detection method comprises combining influenza virus and the detection composition in the environment with the pH value of 7.0-7.2, and then detecting the firefly luciferin signal. According to the detection composition contained by the real-time detection kit, by limiting the ingredients in the detection composition and further limiting the content of the ingredients, detection on ceramidease of influenza virus does not need dividing into two steps, and a one-step reaction can be performed only by directly mixing the detection composition and influenza virus. When relatively high detection sensitivity is guaranteed, operation steps are simplified and real-time detection is realized.

Description

technical field [0001] The invention relates to the technical field of influenza virus detection, in particular to a real-time detection kit and method for influenza virus. Background technique [0002] Influenza is a persistent and serious threat to public health. With the development of medical science, several drugs are currently used to treat or prevent influenza A and B. However, the premise of effective prevention, control or treatment of influenza lies in the rapid, sensitive and specific detection of influenza virus in the early stage of individual infection or large-scale outbreak. [0003] Traditional methods of influenza virus detection usually first require the culture of nasal washes, throat swab samples, airway puncture to aspirate secretions, bronchial lavage fluid or lung tissue, and detect the influenza virus by hemadsorption test on the 2-5th day after culture. Viruses are cultured for detection and generally followed by immunofluorescence assay (IFA) wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/66
CPCC12Q1/66C12Q1/04
Inventor 李兴祥
Owner CELLEX BIOLOGICAL TECH SUZHOU CO LTD
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