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Hemolytic streptococcus diagnostic immunochromato reagent, kit, and detection method

By using a combination of cyclic oligosaccharides and nitrite in an immunochromatography kit to generate nitrous acid and stabilize pH conditions, the problems of low sensitivity and high false positive rate in the detection of hemolytic streptococci in the existing technology are solved. Efficient, fast and accurate detection results are achieved.

Inactive Publication Date: 2016-03-30
TANAKA KIKINZOKU KOGYO KK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

), sometimes depending on the measurement sample, measurement environment, and measurement conditions, the aggregation of the insoluble carrier may still occur, causing non-specific reactions, and problems such as slow development speed may not be solved.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0175] (1) Fabrication of the judgment section on the chromatography medium (1)

[0176] A sheet made of nitrocellulose (manufactured by Millipore, trade name: HF120, 250 mm×25 mm) was used as a film. The rabbit-derived anti-hemolytic streptococcus polyclonal antibody (primary antibody) was diluted with 10 mM phosphate buffer (pH 7.4) containing 5% by mass of isopropanol to a concentration of 1.0 mg / ml, 150 μL of this diluted solution was applied to a membrane with a width of 1 mm using an antibody coater (manufactured by BioDot Co., Ltd.), dried at 50° C. for 30 minutes, and dried overnight at room temperature, and thus deposited on the chromatography medium (1). Create a judgment section.

[0177] (2) Preparation of labeled substance solution

[0178] Anti-hemolytic streptococcus derived from rabbits diluted to a concentration of 0.1 mg / mL with phosphate buffer (pH 7.4) was added to 0.5 mL of gold colloidal suspension (manufactured by Tanaka Kikinzoku Kogyo Co., Ltd.: aver...

Embodiment 4～12

[0201] Sodium nitrite, cyclodextrin, and citric acid were set to various amounts described in Table 2 for each test (test), and implemented in the same procedure form as in Example 1. For positive specimens, use 150 μL concentration of 2 × 10 in each test (test). 6 org / mL of inactivated hemolytic streptococci.

[0202] Then, it judged according to the same visual judgment standard. The judgment results are shown in Table 2.

Embodiment 13

[0204] The procedure was carried out in the same manner as in Example 1 except that tartaric acid was contained in the antigen extraction part (3) instead of citric acid. Then, it judged according to the same visual judgment standard. The judgment results are shown in Table 2.

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PUM

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Abstract

Provided is an immunoassay technique in which an antigen of a target to be detected in an analyte is extracted by an extraction agent, wherein the immunoassay technique is characterized by being carried out in the presence of a cyclic oligosaccharide. The technique is also characterized by the use of an immunochromato kit comprising: an analyte diluent; an immunochromatography medium having an analyte dilution liquid, a specimen dripping part, an antigen extracting part, a marker substance retaining part, and a detection part; and an absorbing part, and used to detect gram-positive bacteria in an analyte in which an organic acid is retained in the antigen extracting part, wherein the immunochromato kit includes a cyclic oligosaccharide and a nitrite compound, through inclusion of any one or more of cyclic oligosaccharides, nitrite compounds, and mixtures of these in either the analyte dilution liquid and / or the specimen dripping part, or both.

Description

technical field [0001] The present invention relates to an immunochromatographic reagent of high importance as a simple in vitro diagnostic kit or a portable diagnostic device for detecting Gram-positive bacteria, especially hemolytic streptococci, in a sample liquid A cartridge, a reagent composition or a sample processing solution used therein, and a detection method. In particular, it relates to a detection method that does not need to prepare the nitrous acid required for the extraction of polysaccharides required for the detection of hemolytic streptococci in each pass, but generates it on the detection equipment, thereby improving the detection efficiency and detection accuracy. method. Background technique [0002] In recent years, with regard to strip-form immunoassays for immunochromatography, the use of simple in vitro diagnostic kits or portable diagnostic devices for detecting analytes (antigens, etc.) in sample fluids by utilizing the specific reactivity of ant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543
CPCG01N2400/18G01N33/5306G01N33/54393G01N33/56944G01N2400/10G01N33/54388G01N2469/10
Owner TANAKA KIKINZOKU KOGYO KK