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Method for producing and using trauma vibrio fast detection kit

A detection kit and rapid technology, applied in biochemical equipment and methods, microbial-based methods, microbial measurement/inspection, etc., can solve the problems of cumbersome operation, time-consuming, and inability to meet the needs of disease prevention and control, and achieve high sensitivity , strong specific effect

Inactive Publication Date: 2008-08-20
INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented quick test system uses special primer sets with unique base regions on certain genes found within Vulna bacteria called Glu A. These areas share these bases with other viruses like Yellow River virus and Lassa fever. They also have similar DNA structures around them. By comparing this data between existing methods such as ELISA and Western blots it becomes possible to identify any type of vibroetta niqueacea without requiring costly equipment.

Problems solved by technology

This patented technical problem addressed in this patents relates to identifying specific types of microorganisms called vibratorella spiroformis during acute illnesses like cholera, causing serious complications including inflammatory syndrome, hemorrhagism, pulmonary embolism, urinary tract obstruction, abdominal pain, menstruation bleeding, dermal damage, and reduced survival rates among patients who contract these diseases through exposure to contaminating water sources overlooking them without any signs until afterward.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Make vibrio vulnificus rapid detection kit by following formula and comprise following (1)-(4):

[0037] (1) LAMP reaction solution A:

[0038]Including 2.5 μL 10× Thermopol reaction buffer, 1.0 μL 10 mmol / L dNTP, 1.0 μL 20 μmol / L upstream internal primer (FIP), 1.0 μL 20 μmol / L downstream internal primer (BIP), 0.25 μL 20 μmol / L upstream external primer (F3), 0.25 μL 20 μmol / L downstream outer primer (B3), 0.5 μL 100 mmol / L MgSO4, 12.5 μL 2mol / L betaine, and 4 μL ddH2O (sterilized double distilled water).

[0039] The upstream internal primer 5-ACCTGTGTTCCATGCTGCTTCT-ATTCGCAACTGAC ATGTCGG-3 described therein;

[0040] Downstream inner primer 5-TAACAAAGGTCACCGTCCAGGC-CAGAACGCAGGTCTTGTG AA-3;

[0041] Upstream outer primer 5-CCAGAGCCAGAGTTCTTCCT-3;

[0042] Downstream outer primer 5-GGCCCATCTCTTCCATCAC-3.

[0043] The 10× Thermopol reaction buffer described therein contains trihydroxymethylaminomethane-hydrochloric acid (Tris-HCl) of 200mmol / L pH8.8, 100mmol / L potass...

Embodiment 2

[0061] Make vibrio vulnificus rapid detection kit by following formula and comprise following (1)-(4):

[0062] (1) LAMP reaction solution A:

[0063] Including 2.5 μL 10× Thermopol reaction buffer, 1.0 μL 10 mmol / L dNTP, 1.0 μL 20 μmol / L upstream internal primer (FIP), 1.0 μL 20 μmol / L downstream internal primer (BIP), 0.25 μL 20 μmol / L upstream external primer (F3), 0.25 μL 20 μmol / L downstream outer primer (B3), 0.5 μ / L 100 mmol / L MgSO 4 , 12.5 μL 2mol / L betaine and 4 μL ddH 2 O (sterilized double distilled water).

[0064] The upstream internal primer, downstream internal primer, upstream external primer, and downstream external primer are the same as above.

[0065] The 10× Thermopol reaction buffer described therein contains trihydroxymethylaminomethane-hydrochloric acid (Tris-HCl) of 200mmol pH8.8, 100mmol / L potassium chloride (KCl), 100mmol / L ammonium sulfate ((NH 4 )2SO 4 ), 20mmol / L magnesium sulfate (MgSO 4 ) and 1% Triton X-100 (Triton X-100);

[0066] (2) U...

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Abstract

The invention relates to a preparation and using method of a vibrio vulnificus rapid detection kit; wherein, the interior of the kit includes a loop-mediated isothermal amplification reaction liquid A, a Bst DNA polymerase B and a chromogenic agent C; the reaction liquid A contains a reaction buffer liquid, a dNTP, a magnesium sulfate, an upstream internal primer 5-ACCTGTGTTCCATGCTGCTTCTATTCGCAACTGACATGTCGG-3, a downstream internal primer 5-TAACAAAGGTCACCGTCCAGGCCAGAACGCAGGTCTTGTGAA-3, an upstream external primer 5-CCAGAGCCAGAGTTCTTCCT-3, a downstream external primer 5-GGCCCATCTCTTCCATCAC-3 and a betaine; the method for detecting the vibrio vulnificus includes the DNA extraction of the samples or the bacteria to be detected, the loop-mediated isothermal amplification reaction of the vibrio vulnificus and the chromogenic detection. The preparation and using method has the advantages of swiftness, strong specificity, high sensitivity and low cost.

Description

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Claims

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Application Information

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Owner INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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