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Detection and detecting kit of enterorrhagia Bacillus coil 0157 nucleic acid

A technology for Escherichia coli and enterohemorrhagic Escherichia coli, which is applied in the detection field of enterohemorrhagic Escherichia coli, can solve the problems of difficulty in adapting to diagnosis, treatment, poor specificity and low sensitivity, and achieves the effects of fast identification method, simple operation and high sensitivity

Inactive Publication Date: 2005-10-12
XIAMEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide an E.coli 0157 detection method that is slow in identification, low in sensitivity, poor in specificity, and difficult to adapt to the problems of diagnosis and treatment during epidemics, and provides a method that is easy to operate, high in experimental reliability, capable of real-time detection, and high in sensitivity. Enterohaemorrhagic Escherichia coli 0157 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
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Effect test

Embodiment 1

[0050] Example 1: Real-time fluorescent PCR detection of isometric fluorescent probe pair 0157

[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] Example 2: Detection of isometric fluorescent probes to 0157 from different sources (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 pair 0157, Shigella, invasive Escherichia coli (EIEC), the detection of Salmonella

[0055] Take 0157, 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 the detection of enterohemorrhagic Escherichia coli, in particular to a method for detecting enterohemorrhagic Escherichia coli 0157 by using the equal-length double-strand fluorescent probe technology and a detection kit thereof. Background technique [0002] Enterohaemorrhagic Escherichia coli 0157 (E.coli 0157) 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 0157 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 Applications(China)
IPC IPC(8): C12Q1/04C12Q1/68
Inventor 陈亮于广福邵寒娟牛建军
Owner XIAMEN UNIV
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