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Detection and detecting kit of enterorrhagia Bacillus coil 0157 nucleic acid

An Escherichia coli and enterohemorrhagic technology, applied in the detection field of enterohemorrhagic Escherichia coli, can solve the problems of difficult to adapt to diagnosis, treatment, low sensitivity, poor specificity, etc., and achieve the effects of fast identification method, high sensitivity and easy operation.

Inactive Publication Date: 2008-11-26
XIAMEN UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

[0006] The present invention aims at the current E.coli O157 detection method with slow identification, low sensitivity, poor specificity, difficult to adapt to the problems of diagnosis and treatment during the epidemic, and provides a method with simple operation, high experimental reliability, real-time detection and high sensitivity. Enterohaemorrhagic Escherichia coli O157 nucleic acid detection method and its kit with good specificity and short cycle

Method used

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  • Detection and detecting kit of enterorrhagia Bacillus coil 0157 nucleic acid
  • Detection and detecting kit of enterorrhagia Bacillus coil 0157 nucleic acid
  • Detection and detecting kit of enterorrhagia Bacillus coil 0157 nucleic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1: Real-time fluorescent PCR detection of O157 by equal-length fluorescent probes

[0051] Take 1 μl of the sample and add it to the detection solution Mixs of the detection kit, the final volume is 20 μl. The reaction program of real-time fluorescent PCR is that the PCR amplification program is denaturation at 94°C for 3 minutes, entering cycle, denaturation at 94°C for 15 s, annealing at 55°C for 20 s, extension at 72°C for 20 s, 40 cycles, [Gain Fam]=9. Fluorescent signal intensities were collected during the annealing phase.

Embodiment 2

[0052] Embodiment 2: Detection of different sources of O157 by equal-length fluorescent probes (sample originates from polluted water, food, human and animal feces, etc.)

[0053] Take 1 μl of each sample and add it into the detection solution, and perform real-time fluorescent PCR reaction according to the method of the implementation procedure. The fluorescence intensity of different samples rises at different heights, which represents a positive result. (see figure 1 )

Embodiment 3

[0054] Embodiment 3: Isometric fluorescent probe pairs O157, Shigella, invasive Escherichia coli (EIEC), the detection of Salmonella

[0055] Take O157, Shigella, invasive Escherichia coli (EIEC), and Salmonella samples 1 μl each into the detection solution, and perform real-time fluorescent PCR reaction according to the method of the implementation procedure. If the fluorescence intensity rises, it represents a positive result. ( see figure 2 )

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Abstract

A nucleic acid detection method and diagnostic kit for intestinal hemorrhage bacillus coli O157. It adopts equal-length double-chain fluorogence probe technology. It is credible with high sensitivity and good specificity. The detection can be real-time operated with short periodicity.

Description

technical field [0001] The invention relates to a detection of enterohaemorrhagic Escherichia coli, in particular to a method for detecting enterohaemorrhagic Escherichia coli O157 by adopting equal-length double-strand fluorescent probe technology and a detection kit thereof. Background technique [0002] Enterohemorrhagic Escherichia coli O157 (E.coli O157) is a major serotype of diarrhea-causing Escherichia coli, which can cause hemorrhagic enteritis, accompanied by severe abdominal pain, fever, and severe renal insufficiency, hemolytic Uremic syndrome (HUS), hemolytic anemia, thrombocytopenic purpura, and even death due to acute and chronic renal failure. Food poisoning caused by E.coli O157 has occurred one after another in developed countries such as the United States, Japan, the United Kingdom, Canada, and Australia. In 1993, an outbreak involving more than 700 children in 4 states occurred in the United States, of which 51 cases developed hemolytic uremic syndrome (...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 陈亮于广福邵寒娟牛建军
Owner XIAMEN UNIV
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