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Immunochromatography-assisted detection method

An immunochromatographic detection, immunochromatographic technology, applied in immunoassays, measuring devices, analytical materials, etc., can solve the problem of unknown methods to eliminate this problem, low detection sensitivity of influenza B virus, unknown elimination of such bleaching phenomena, methods, etc., to achieve the effect of accurate measurement

Active Publication Date: 2016-07-20
SEKISUI MEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no known way to eliminate this bleaching phenomenon
[0007] On the other hand, although immunochromatography has been put into practical use in a large number of rapid clinical tests (point-of-care tests) and is particularly widely used in the diagnosis of infectious diseases (such as influenza), such as using other immunoassays (such as LTIA ), immunochromatography has a problem of low detection sensitivity for influenza B virus, and a method for eliminating this problem is not yet known

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0115] Example 1 (bleaching evaluation test)

[0116] Bleaching was evaluated by using an immunochromatographic test strip (manufactured by Sekisui Medical Co., Ltd., Rapid Testa (registered trademark) ColorFLUStick, Lot. K1221221077P) stored unopened for two years at the temperature described in the package insert. Add glycerol or methanol to the sample-diluent (Tris buffer (pH 8.5)) attached to the test strip, respectively, to prepare sample dilutions containing 1%, 5% and 10% glycerol or 1%, Sample dilutions of 5% and 10% methanol. A set (n=3) of immunochromatographic test strips was immersed in each sample dilution to allow the sample dilution to run on the chromatograph for 10 minutes. Then, each test strip was visually inspected for the occurrence of bleaching. The results are shown in Table 1.

[0117] Similarly, the same test was performed submerging the test strips in sample dilutions containing 1%, 5% and 10% ethanol. The results are shown in Table 2.

[0118] "...

Embodiment 2

[0127] Embodiment 2 (accelerated test)

[0128] The immunochromatographic test strips obtained immediately after preparation and the immunochromatographic test strips stored at 60° C. for 10 days and 20 days were immersed in sample diluents containing 5% and 10% methanol to allow the solutions to be as described in Example 1. The situation was developed for 10 minutes and the occurrence of bleaching was checked visually. The results are shown in Table 3.

[0129] [table 3]

[0130]

[0131] d: number of days

[0132] (result)

[0133] Inhibition of the occurrence of bleaching and a reduction in the extent of bleaching was observed in all immunochromatographic test strips used immediately after preparation and after storage at 60°C for 10 and 20 days when sample diluents containing 5% and 10% methanol were used . In particular, a significant inhibition of the occurrence of bleaching was observed when using a sample diluent containing 10% methanol.

Embodiment 3

[0134] Embodiment 3 (comparison of tinting strength in A line and B line)

[0135] Compare the intensity of staining in "Line A" and "Line B" using immunochromatographic test strips. Prepare samples by mixing 330 μL of sample diluent containing 5% and 10% methanol with 50 μL of sample inactivated by diluting influenza A and influenza B virus strains at the dilution factors described in Tables 4 and 5 Viral antigens (respective types of influenza viruses are described in the table below). A set (n=2) of test strips was submerged in 135 [mu]L of the sample to which the respective inactivated viral antigen was added, allowing the sample dilutions to spread on the test strips for 10 minutes. Then, based on the color samples, the coloring intensity at the detection portion of each virus was obtained. The results for influenza A virus and influenza B virus are shown in Table 4 and Table 5, respectively.

[0136] [Table 4]

[0137]

[0138] Conv.: Conventional

[0139] Meth.:...

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Abstract

The present invention provides a detection method using a immunochromatography test strip, said method exhibiting high accuracy and sensitivity while minimizing the incidence of "white streaking." The present invention provides a immunochromatography-assisted detection method involving the use of an immunochromatography test strip. The immunochromatography test strip has: (1) a conjugate pad having a sample supply portion for supplying a sample that can contain a substance to be detected; and a conjugate portion containing a conjugate in which an antibody for the substance to be detected is immobilized on a marker, downstream from the sample supply portion; and (2) an insoluble membrane carrier having at least one detecting portion on which the antibody for the substance to be detected is immobilized. The detection method includes (A) a step for supplying to the sample supply portion methanol and a sample that can contain the substance to be detected, and (B) a step for detecting the substance to be detected as an immunoreaction product on the insoluble membrane carrier.

Description

technical field [0001] The invention relates to an immunochromatographic detection method. The invention also relates to a method for detecting influenza B virus. Background technique [0002] A detection method using an immunochromatographic test strip is known as a method of detecting an analyte (substance to be detected) in a sample by an antigen-antibody reaction. Immunochromatography is a method that involves combining an analyte with a labeled conjugate (the labeled conjugate may also be referred to as a conjugate) in which an antibody to the analyte is immobilized. The immune complex is developed together with a mobile phase (such as a buffer) through a stationary phase, which is an insoluble membrane support having a detection portion on which antibodies against the analyte are immobilized as A capture agent, whereby immune complexes captured by the capture agent are detected. Colloidal metal particles, such as colloidal gold, and colored latex particles, are used...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/531G01N33/576
CPCG01N33/5761G01N33/54388G01N33/56983G01N2333/11G01N2469/10
Inventor 西谷公良
Owner SEKISUI MEDICAL CO LTD
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