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Special probe and gene chip used for identifying pathogenic aspergillus

A pathogenic and Aspergillus technology, which is applied in the field of special probes and gene chips for identifying pathogenic Aspergillus, and can solve problems such as inability to specifically detect pathogenic Aspergillus

Inactive Publication Date: 2013-06-26
PEKING UNIV FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is still not possible to quickly and specifically detect pathogenic Aspergillus in clinical specimens and further identify the species

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1. Detection of pathogenic Aspergillus and rapid identification of strains

[0044] Extract the genomic DNA from the samples containing the pathogenic Aspergillus of different known strains according to the standard method, and then use the above primer pair (sequence 11 and 12 in the sequence list) to perform PCR reaction (about 1.5 hours), and then the PCR product With the nitrocellulose membranes labeled with different probes (respectively the probes shown in sequence 1 to sequence 10), the flow-through hybridization reaction is carried out through the medical nucleic acid molecular hybridization instrument, and the bacterial species in the tested sample are judged according to the result of the reaction. The time required is 3.5 hours. The results are shown in Table 1.

[0045] Table 1 Identification results of common pathogenic Aspergillus

[0046] Sample

Embodiment 2

[0047] Example 2: Detection of pathogenic Aspergillus mixed bacteria and rapid identification of strains

[0048] A sample containing a mixture of known pathogenic Aspergillus strains was extracted according to standard methods to extract genomic DNA, and then the primer pair (sequence 11 and 12 in the sequence listing) was used for PCR reaction (about 1.5 hours), and then the PCR product was combined with The nitrocellulose membranes labeled with different probes (respectively the probes shown in sequence 1 to sequence 10) are subjected to a flow-through hybridization reaction (about 2 hours) through a medical nucleic acid molecular hybridization instrument, and the test sample can be judged according to the result of the reaction The total time required for the mixed strains is 3.5 hours. The results are shown in Table 2.

[0049] Table 2 Identification results of mixed samples of pathogenic Aspergillus

[0050] Specimen

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PUM

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Abstract

The invention discloses a special probe and gene chip used for identifying pathogenic aspergillus. The special probe for identifying pathogenic aspergillus provided by the invention comprises at least one of the following 20 DNA fragments: 10 DNA fragments of which nucleotide sequences are from sequence 1 to sequence 10 in the sequence table and 10 DNA fragments of which nucleotide sequences are complementary sequences of the sequence 1 to sequence 10 in the sequence table. The invention also provides a DNA chip containing the special probe for identifying pathogenic aspergillus. The special probe, DNA chip or device is all used for identifying pathogenic aspergillus. When in use, the special probe is coated on a low-density gene chip membrane, then the flow-through hybridization between the ITS region of rDNA of the pathogenic aspergillus to be tested and the species specificity probe on the low-density gene chip membrane is carried out by using the test platform based on the flow-through hybridization technology, and then color development is carried to finally identify the pathogenic aspergillus. The special probe and gene chip of the invention are characterized by strong specificity and fast operation and can play fast, specific and positive role in testing the common pathogenic aspergillus in clinical specimens and identifying the strain of the pathogenic aspergillus.

Description

Technical field [0001] The invention relates to a special probe and gene chip for identifying pathogenic Aspergillus. Background technique [0002] In recent years, with the in-depth development of anti-malignant tumor therapy, organ transplantation, especially the continuous implementation of allogeneic hematopoietic stem cell transplantation (HSCT), invasive aspergillosis (Invasive aspergillosis, IA) caused by Aspergillus infection has become a variety of severe immunity The most common opportunistic infection in the damaged body is also the leading cause of patient death. For example, the fatality rate in leukemia and HSCT recipients often exceeds 90%. The pathogens that cause IA include Aspergillus fumigatus, Aspergillus flavus, Aspergillus nidulans, Aspergillus terreus, Aspergillus niger, etc. The susceptibility of these IA pathogens to different antifungal drugs is very different. For example, Aspergillus fumigatus is sensitive to amphotericin B, but Aspergillus terreus is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 刘伟许辉李若瑜
Owner PEKING UNIV FIRST HOSPITAL