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Oligonucleotide probe kit for detecting common intestine trac kpathogenic bacteria and its use

An oligonucleotide probe, oligonucleotide technology, applied in the determination/inspection of microorganisms, resistance to vector-borne diseases, biochemical equipment and methods, etc., can solve problems such as inability to achieve

Inactive Publication Date: 2005-10-19
RADIOLOGY INST ACAD OF MILITARY MEDICINE SCI PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the simultaneous identification of a large number of intestinal pathogenic bacteria, the current commonly used methods cannot meet the requirements

Method used

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  • Oligonucleotide probe kit for detecting common intestine trac kpathogenic bacteria and its use
  • Oligonucleotide probe kit for detecting common intestine trac kpathogenic bacteria and its use
  • Oligonucleotide probe kit for detecting common intestine trac kpathogenic bacteria and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1. Screening and optimization of oligonucleotide probes for detection of seventeen species of enteropathogenic bacteria

[0044] 1. Design of oligonucleotide probes for detection of seventeen species of enteropathogenic bacteria

[0045] The probes were designed on the bacterial 16S rRNA and 23S rRNA and the large plasmid in Shigella dysenteriae encoding invasion plasmid antigen H (ipaH) and Salmonella typhi regulating the expression of Vi antigen and other gene sequences, with high sensitivity and specificity, oligonucleotides The sequence of the acid probe is consistent with the base sequence of the positive strand of DNA in the specific region of the above-mentioned gene (see Table 1-1 for the sequence of the probe). The probe length is between 25-50 bases. When the probe is synthesized, its 3' terminal amino group is modified, which is used for the probe to react with the active aldehyde group on the surface of the slide and fix it to the surface of the car...

Embodiment 2

[0185] Example 2. Specificity and Sensitivity Evaluation of Oligonucleotide Probes

[0186] 1. Gene chip preparation Dilute the oligonucleotide probes with spotting solution (6×SSC, 0.1% SDS) to a final concentration of 50 μmol / L, and transfer 5ul to a 384-well plate. The probes were spotted on the aldehyde-based chip with a Cartesian chip preparation instrument (see Figure 4 for the arrangement of the probes, and see Figure 5 for the hybridization results of the chips with fluorescently labeled PCR products of seventeen species of enteropathogenic bacteria). In the probe microarray, the reverse complementary sequence of the fluorescent primer is used as a positive control, the irrelevant random sequence is used as a negative probe control, and the blank sample solution without probe is used as a negative control. The probe sequences are shown in Table 2-1.

[0187] Table 2-1 Gene chip probe sequences for detection of common intestinal pathogens

[0188] Pr...

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Abstract

The present invention belongs to the field of microbe detecting technology. The oligonucleotide probe for detecting common intestinal tract pathogenic bacteria is designed on 16S rRNA and 23S rRNA of bacteria, ipaH of dysentery bacillus giant plasmid, VipR of Salmonella typhi and other gene sequence, has length of 25-50 bp, and relatively high sensitivity and specificity. The oligonucleotide probe is suitable for detection based on nucleic acid hybridization principle, especially detection based on gene chip principle. Under certain use condition, it can detect Listeria, parahemolutic vibrio, Campylobacter, etc. It may be used in many aspects, such as disease diagnosis, environment detection, food poisoning detection, etc.

Description

technical field [0001] The invention belongs to the technical category of microorganism detection technology, especially the nucleic acid detection technology of intestinal pathogenic bacteria. The invention relates to a set capable of detecting Listeria, Vibrio parahaemolyticus, Campylobacter, Yersinia enterocolitica, Yersinia, Staphylococcus aureus, Clostridium botulinum, wax sample Oligonucleotide detection of Bacillus, Salmonella, Salmonella typhi, Shigella, Shigella, Clostridium perfringens, Proteus, Brucella, EHE O157:H7 and Vibrio cholerae needle, the sequence and use of the set of probes. Background technique [0002] Enteropathogenic bacteria are the main pathogens that cause infectious diseases with symptoms of diarrhea and vomiting clinically. Common pathogens include Listeria, Vibrio parahaemolyticus, Campylobacter, Yersinia enterocolitica, Yersinia spp, Staphylococcus aureus, Clostridium botulinum, Bacillus cereus 17 species, including Salmonella, Salmonella ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCY02A50/30
Inventor 王升启金大智文思远陈苏红
Owner RADIOLOGY INST ACAD OF MILITARY MEDICINE SCI PLA
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