Low-noise excitation type fluorescent marker based immunochromatography test strip for vibrio parahaemolyticus

An immunochromatographic test strip and Vibrio hemolyticus technology, which is applied in the field of medical detection, can solve the problems of difficult and rapid quantitative detection of Vibrio parahaemolyticus, etc., and achieves the effects of improved sensitivity and high detection sensitivity

Inactive Publication Date: 2015-04-01
BEIJING ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECH CENT +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] For this reason, the technical problem to be solved by the present invention is the problem that it is difficult to rapidly quantitatively detect Vibrio parahaemolyticus in the prior art, and then provide a kind of Vibrio parahaemolyticus immunochromatographic test strip based on low-noise excitation fluorescent labeling, and It further discloses its application

Method used

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  • Low-noise excitation type fluorescent marker based immunochromatography test strip for vibrio parahaemolyticus
  • Low-noise excitation type fluorescent marker based immunochromatography test strip for vibrio parahaemolyticus
  • Low-noise excitation type fluorescent marker based immunochromatography test strip for vibrio parahaemolyticus

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Embodiment 1

[0042] The Vibrio parahaemolyticus immunochromatographic test strip based on low-noise excitation fluorescent label described in this embodiment is as follows: figure 1 As shown, it includes a bottom plate 7 and a sample pad 1, a binding pad 2, an antibody-carrying membrane 3 and a water-absorbent pad 6 that are sequentially adhered to the bottom plate along the length direction of the bottom plate 7; and the sample pad 1 , binding pad 2, antibody-carrying membrane 3 and water-absorbing pad 6, which are sequentially in contact with and only partially overlap adjacent parts; the antibody-carrying membrane 3 is adhered to the middle part of the bottom plate 7, and A quality control line 5 with spaced detection lines 4 is provided, the detection line 4 is close to the binding pad 2, and the quality control line 5 is close to the water-absorbing pad 6; the quality control line 4 and the detection line 5 are arranged in parallel, and the distance between the detection line and the ...

Embodiment 2

[0052] This embodiment provides a method for qualitative detection of Vibrio parahaemolyticus using the low-noise excitation-based fluorescence-labeled Vibrio parahaemolyticus immunochromatographic test strip, including the following steps:

[0053] a) Take 25g of fish sample, cut it into pieces, dilute the sample with 225mL PBS (pH 7.4) buffer solution containing 5% BSA, 0.1% Tween 20, take the eluate for ultrasonic destruction, probe with a diameter of 3mm, work for 5s, rest for 5s , 40% energy (the ultrasonic cell pulverizer provided by Ningbo Xianchang Electronic Technology Co., Ltd., the model is XC-CD type), a total of 30 cycles, draw 50 μL of the ultrasonically treated test sample and drop it on the sample pad 1, wait After the absorption is dry, 50 μL of chromatographic solution is added dropwise, and the buffer solution is taken as a negative control, and the test strip is used for immunochromatography;

[0054] b) After standing at room temperature for 15 minutes, us...

Embodiment 3

[0057] The method for the quantitative detection of Vibrio parahaemolyticus using the described low-noise excitation-based fluorescent-labeled Vibrio parahaemolyticus immunochromatographic test strip:

[0058] Preparation of standard curve: take 10 7 CFU / mL Vibrio parahaemolyticus pure bacteria (isolated strains in this laboratory), the pure bacteria were carried out multiple serial dilutions with PBS (pH7.4) diluent to make 2.4*10 5 CFU / mL, 1.2*10 5 CFU / mL, 2.4*10 4 CFU / mL, 1.2*10 4 CFU / mL, 2.4*10 3 CFU / mL, 1.2*10 3 CFU / mL, 2.4×10 2 CFU / mL, 1.2×10 2 CFU / mL sample; Ultrasonic bacteriostasis, probe with a diameter of 3mm, work for 5s, rest for 5s, 40% energy (ultrasonic cell pulverizer provided by Ningbo Xianchang Electronic Technology Co., Ltd., model XC-CD), a total of 30 times Circulate, absorb 50 μL of the treated sample and drop it on the sample pad 1, then add 50 μL of the above PBS buffer solution dropwise after the absorption is dry; let stand at room temperature...

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Abstract

The invention relates to a low-noise excitation type fluorescent marker based immunochromatography test strip for vibrio parahaemolyticus as well as a preparation method and an application of the immunochromatography test strip. A low-noise excitation type fluorescent dye is adopted as a marker, and an immunochromatographic technology is adopted, so that the immunochromatography test strip targeting vibrio parahaemolyticus is prepared; during detecting, a special portable low-noise excitation type fluorescence scanner is adopted to scan a quality control line and a detection line respectively so as to realize qualitative and quantitative detection of a sample according to a fluorescence intensity detection value of the detection line; and the test strip is suitable for instant detection for vibrio parahaemolyticus in food and pathological samples and has the characteristics of fastness, high sensitivity, consideration of both qualitative and accurately quantitative detection and portability.

Description

technical field [0001] The invention belongs to the field of medical detection, and in particular relates to a Vibrio parahaemolyticus immunochromatographic test strip based on a low-noise excitation type fluorescent dye label and a preparation method and application thereof. Background technique [0002] As an important indicator for the detection of pathogenic bacteria, Vibrio parahaemolyticus is of great significance in public health, food hygiene, animal husbandry and veterinary medicine, and entry-exit inspection and quarantine. Detection puts forward higher requirements. In order to seek fast, accurate, high-sensitivity, easy-to-operate and quantitative detection methods, scholars from all over the world have conducted a lot of research, developing from traditional methods to rapid detection methods based on immunology and molecular biology, and in practice Continuously making new progress. [0003] Immunochromatography technology is a new type of immunoassay technol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/533G01N21/64
CPCG01N33/56911G01N21/6402G01N33/533
Inventor 张捷杨向莹许美玲邹明强汪琦张晓光舒咬根陈广全
Owner BEIJING ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECH CENT
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