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Wound vibrio quantitative detecting test strip

A technology for quantitative detection of Vibrio vulnificus, which is applied in the determination/inspection of microorganisms, fluorescence/phosphorescence, resistance to vector-borne diseases, etc. Fast, stable results

Inactive Publication Date: 2010-09-01
NINGBO BOAO BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The traditional method goes through multiple screening steps, requires a variety of media, and takes a long time; there is another method using vvh monoclonal antibody detection. However, the time required for the detection of both methods is relatively long. In order to minimize human contact with or consumption of aquatic products contaminated by Vibrio vulnificus, it is extremely important to establish a sensitive and specific detection method to protect human health.

Method used

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Embodiment Construction

[0038] Below in conjunction with accompanying drawing embodiment the utility model is described in further detail:

[0039] (1) Sample processing and template extraction

[0040] Take aquatic products for high-speed homogenization of dehulled tissues, take homogenate enrichment solution for 12 hours, take 1.5Ml enrichment solution and centrifuge at 10000rpm for 5min, discard the supernatant, add 30μL double distilled water and boil for 5min, centrifuge at 12000rpm for 10min, then take the supernatant 2 μL was used as template DNA to be detected.

[0041] (2) Amplification of the target fragment of Bacillus vulnificus

[0042] Take the amplification reaction solution, add reaction buffer, vvhA1, vvhA2, template, enzyme, and finally double-distilled water in sequence to form a reaction system with a total volume of 20-100 μL, mix well, and centrifuge briefly. The reaction program is 94°C for 5 minutes, 94°C 30 cycles of 30s, 55°C for 30s, 72°C for 30s, 72°C for 6min, and final...

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Abstract

The invention discloses a primer, a probe, a wound vibrio detecting method and a detecting kit for wound vibrio detection in aquatic products. The target gene of the primer and probe combined site is vvhA350 site-800 site of the cytolysin structure gene of the wound vibrio. The wound vibrio in the aquatic products is quantitatively detected by a wound vibrio quantitative detecting test strip, comprising detecting test paper and reaction buffer solution, wherein the detecting test paper consists of a sample absorbing zone, a result scanning window and a terminal indicating window. The invention is characterized in that the marking object is the epitropous phosphor particle, has the advantages of quantitative detection, good sensitivity, stability and safety, and can quantitatively detect the wound vibrio in the aquatic products.

Description

technical field [0001] The present invention relates to a test strip for quantitative detection of Vibrio vulnificus gene diagnosis in aquatic products based on up-conversion phosphorescence (UPT) nucleic acid chromatography technology, in particular to primers and probes specific to specific gene segments of Vibrio vulnificus; The invention relates to an up-conversion phosphorescent nucleic acid chromatography detection method and a detection kit for detecting Vibrio vulnificus in aquatic products by using the above primers and probes with a PCR amplification method. This detection method is used for sensitive, rapid and quantitative detection of Vibrio vulnificus in aquatic products. Background technique [0002] Vibrio vulnificus is a halophilic pathogenic bacteria widely distributed in oceans and salt lakes. This bacteria is very harmful to patients with diabetes, alcoholic liver disease, liver cirrhosis, hepatitis and unexplained liver dysfunction, AIDS or other serious...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06G01N21/64
CPCY02A50/30
Inventor 范建忠范翠翠黄宝福宋超霞游玉容
Owner NINGBO BOAO BIOENG
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