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Kit and method for detection of staphylococcus aureus by fine-pitch array electrode based immune quantitative sensor

A staphylococcus, immune quantitative technology, applied in the biological field, can solve the problems of complex and cumbersome inspection procedures, time-consuming, low detection sensitivity, etc., and achieve the effect of simple operation steps, small background interference, and high sensitivity

Active Publication Date: 2016-03-02
长沙市食品药品检验所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Traditional methods for detecting bacteria include biochemical culture, isolation and identification, the inspection procedures are complex and cumbersome, and it takes about 4-7 days to report the inspection results, which takes too long and the detection sensitivity is low

Method used

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  • Kit and method for detection of staphylococcus aureus by fine-pitch array electrode based immune quantitative sensor
  • Kit and method for detection of staphylococcus aureus by fine-pitch array electrode based immune quantitative sensor
  • Kit and method for detection of staphylococcus aureus by fine-pitch array electrode based immune quantitative sensor

Examples

Experimental program
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Effect test

Embodiment 1

[0038] A kit for the detection of Staphylococcus aureus by an immunoquantitative sensor based on a micro-pitch array electrode. The coated substrate is a polyclonal antibody to Staphylococcus aureus, the primary antibody is a monoclonal antibody to Staphylococcus aureus, and the positive control standard is gold Standard strain of Staphylococcus aureus.

Embodiment 2

[0040]On the basis of Example 1, this embodiment also includes a negative control standard, a blocking solution, a secondary antibody and a substrate solution, the negative control standard is PBS buffer, the blocking solution is a 3% BSA solution, and the secondary antibody It is goat anti-mouse IgG (H+L) labeled with alkaline phosphatase, and the substrate solution is AAP with a concentration of 1mmol / L and AgNO 3 Sterilized glycine-NaOH solution with a concentration of 5 mmol / L.

[0041] Further, the preparation method of PBS buffer solution:

[0042]

[0043] Add ultrapure water to dilute to 1000mL.

[0044] Further, the preparation method of blocking solution: BSA3g, add 100mL sterile 0.01mol / LPBS to prepare.

[0045] Furthermore, the diluent of the primary antibody is a PBS solution containing 0.1% BSA.

[0046] Furthermore, the diluent of the secondary antibody is also included, and the diluent includes the components with the following concentrations: 10mmol / LHEP...

Embodiment 3

[0050] A method for detecting Staphylococcus aureus with an immunoquantitative sensor based on a micro-pitch array electrode using the kit described in Example 1, comprising the following steps:

[0051] (1) Staphylococcus aureus polyclonal antibody diluted with PBS buffer solution, then coated in 96-well micro-pitch array electrode chip wells, 100 μL per well, incubated overnight at 4°C; wash the micro-pitch interdigital array with ultrapure water Electrode three times, add 200 μL of blocking solution to each well of the cleaned electrode, incubate at 37°C for 1 hour, discard the blocking solution, wash with ultrapure water three times, vacuum-dry and plasticize, and store in a refrigerator at 4°C;

[0052] (2) the standard strain concentration of Staphylococcus aureus is 10 8 CFU / mL bacterial solution was heated in a water bath at 100°C for 15 minutes as a positive control standard,

[0053] (3) Take a series of Staphylococcus aureus sample solutions of different concentrat...

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Abstract

The invention discloses a kit and method for detection of staphylococcus aureus by a fine-pitch array electrode based immune quantitative sensor. The coating substrate of the kit is a staphylococcus aureus polyclonal antibody, the primary antibody is a staphylococcus aureus monoclonal antibody, and a positive control standard substance is a staphylococcus aureus standard strain. The kit and the method provided by the invention can complete detection within 3h, have simple operation steps, can finish detection rapidly and accurately, are suitable for batch detection of samples, have strong specificity, high sensitivity and small background interference, can rapidly screen and detect staphylococcus aureus in food, and has wide detection temperature suitable range.

Description

technical field [0001] The invention relates to a kit and a method for detecting Staphylococcus aureus by an immunoquantitative sensor based on a micro-pitch array electrode, belonging to the field of biotechnology. Background technique [0002] Staphylococcus aureus is an important pathogenic bacterium of zoonosis, which widely exists in air, water, dust and human and animal excreta in nature. Both humans and animals have a high rate of bacteria transmission. Healthy people often have enterotoxin-producing (SE) strains in their throats, nasal cavities, skin, and hair, and there are many chances of food being contaminated by them. This year, the US Centers for Disease Control and Prevention reported that infections caused by Staphylococcus aureus accounted for the second place, second only to Escherichia coli. Staphylococcus aureus enterotoxin is a worldwide health problem. In the United States, food poisoning caused by Staphylococcus aureus enterotoxin accounts for 33% of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577
CPCG01N33/56938G01N33/577
Inventor 杨丽霞邱华丽王淑娟聂静苑
Owner 长沙市食品药品检验所
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