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Method of detecting bioterror related pathogen bacteria and its special DNA chip

A technology of DNA chips and pathogenic bacteria, which is applied in biochemical equipment and methods, and the determination/testing of microorganisms, and can solve the problems of unsatisfactory specificity of oligonucleotide probes, weak detection specificity, and cross-reaction phenomena, etc. , to achieve the effects of avoiding non-specific results, high sensitivity and specificity, and avoiding cross-reaction

Inactive Publication Date: 2008-07-23
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to various reasons, the specificity of some oligonucleotide probes designed is not ideal, there is a relatively serious cross-reaction phenomenon, and the detection specificity is not strong

Method used

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  • Method of detecting bioterror related pathogen bacteria and its special DNA chip
  • Method of detecting bioterror related pathogen bacteria and its special DNA chip
  • Method of detecting bioterror related pathogen bacteria and its special DNA chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1. Preparation of a DNA chip containing probes of sequences 1 to 16 in the sequence listing and detection of eight bioterrorism-related pathogenic bacteria

[0076] 1. Preparation of a DNA chip containing probes of sequences 1 to 16 in the sequence listing

[0077] Bacterial Strains: Eight bioterror-related pathogenic bacteria: Bacillus anthracis, Yersinia pestis, Brucella, Francisella tularensis, Burke's pseudomallei Burkholderia pseudomalleii, Rickettsia prowazekii, Rickettsia rickettsii, and Coxiella burnetii; and Yersinia pseudotuberculosis Yersinia pseudotubercolus, Bacillus subtilis, Staphylococcus aureus, Corynebacterium diphtheriae and Salmonella typhi were all purchased from the State Key Laboratory of Biosafety of Pathogenic Microorganisms room. Bacterial template DNA was extracted according to NaI cracking-glass powder adsorption method (Yang Ruifu, Guo Zhaobiao, Zhang Minli, etc. Research on the standardization of nucleic acid diagnostic technology...

Embodiment 2

[0090] Example 2. Experimental verification of eight bioterrorism-related pathogenic bacteria using the chip prepared in the example

[0091] 1. Detection of Yersinia pestis

[0092] 1. Specificity experiment

[0093] Using the DNA of Yersinia pestis, Yersinia pseudotuberculosis, Corynebacterium diphtheriae, Staphylococcus aureus or Salmonella typhi extracted respectively as a template, use the method in Table 2 of (1) of Example 1 step 2 According to the method of (2) of embodiment 1 step 2, each bacterium carries out three groups of multiplex PCR separately respectively, after the amplification reaction ends, the three groups of amplification products of each bacterium are mixed separately respectively, according to the implementation The method of (2) of example 1 step 2 is respectively hybridized with the DNA chip prepared in embodiment 1, and the results are as follows Figure 4 As shown, indicating that the template of Yersinia pestis can generate positive signals at t...

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Abstract

The invention discloses a detecting biological terror correlative pathogenic bacteria method and its special DNA chip. The method steps are using table one primer to do PCR amplification for waited sample DNA; hybridizing the gained amplification product with DNA chip; detecting hybrid signal; if the hybrid signal is positive, the sample contains biological terror correlative pathogenic bacteria. The DNA chip includes at least one probe from sequence one to sixteen. It has good application prospect in fighting biological terror field and clinic microorganism detection.

Description

technical field [0001] The invention relates to a method for detecting pathogenic bacteria related to biological terrorism and a special DNA chip thereof. Background technique [0002] Since the United States experienced the "Anthrax Mail" attack, anti-biological terrorism has become an important part of the international anti-terrorism struggle. One of the prerequisites for dealing with the threat of bioterrorism is the rapid and accurate detection of bioterrorism-related pathogens. [0003] At present, the use of gene chip technology to realize the general detection and identification of bacteria has always been an important field of research by microbiologists at home and abroad. At present, the most commonly used strategy for bacterial universal detection chips is to use the conserved 16S rRNA gene or 23S rRNA gene in bacteria, design universal amplification primers and specific oligonucleotide probes for each bacteria, immobilize on the chip, and use universal primers ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 宋亚军王豫翟俊辉郭兆彪王津杨瑞馥
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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