Gene chip and kit for detecting invasive fungal pathogenic bacteria

An invasive fungal, gene chip technology, applied in the directions of microorganism-based methods, biochemical equipment and methods, and microbial determination/examination, which can solve the problems of difficulty in diagnosing fungal infection, difficulty in samples, false positives, etc., and shorten the detection time. cycle, improving detection sensitivity and specificity, and the effect of high accuracy

Active Publication Date: 2011-02-09
TIANJIN BIOCHIP TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One of these methods is the detection of antigens in serum, however, serum methods to detect C. neoformans antigens have the disadvantage of false positives
Diagnosing fungal infections is often difficult in the laboratory
Culturing samples obtained from patient urine, blood, and saliva is often difficult

Method used

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  • Gene chip and kit for detecting invasive fungal pathogenic bacteria
  • Gene chip and kit for detecting invasive fungal pathogenic bacteria
  • Gene chip and kit for detecting invasive fungal pathogenic bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Design and preparation of embodiment 1 probe

[0036]Ribosomal DNA sequences exist in all organisms, have the same origin and function during evolution, and can reflect comparable evolutionary histories among species. The internal transcribed spacer (Internal Transcribed Spacer, ITS) of rDNA is a region between the 18S and 28S rDNA gene copy regions in eukaryotic rDNA, and the 5.8S rDNA is located between ITS1 and ITS2. The nucleotide sequence of 5.8SrDNA is highly conserved, while ITS1 and ITS2 evolve rapidly, but the inter-species homology is very small, while the intra-species homology is very high, which can be used for inter-species identification.

[0037] 1. Sequence acquisition:

[0038] (1) Acquisition of the 18S-28S region gene sequence: Candida albicans, Candida parapsilosis, Candida tropicalis, Candida kefir, Candida krusei, and Pseudomonas portuguese were downloaded from the GenBank public database. The 18S-28S region gene sequences of Trichomonas, Cand...

Embodiment 2

[0051] Design and preparation of embodiment 2 universal primers

[0052] 1. Universal primer sequence:

[0053]

[0054] 2. Primer synthesis: Entrust the primer synthesis company (Beijing Handsome) to synthesize the primer sequences in the above table, and reserve them for later use.

Embodiment 3

[0055] Example 3 Gene Chip Preparation——Chip Spotting

[0056] 1. Dissolving probes: the probes synthesized in Example 1 were respectively dissolved in 50% DMSO solution, and diluted so that the final concentration of the probes reached 1 μg / μl.

[0057] 2. Adding plate: Add the dissolved probe to the corresponding position of the 384-well plate, 10 μl per well.

[0058] 3. Spotting: as figure 1 The shown 75.5mm×25.5mm×1mm (length×width×height) clean aldehydated glass slide is placed on the stage of the chip spotting instrument (SpotArray 72), and the control software of SpotArray is used to run the program. according to figure 2 The arrangement shown is spotted on the aldehyde-formed glass slide in the spotting area of ​​4.5 mm × 4.5 mm to form a medium-low density DNA micro-array, and the eight dot matrix areas on the glass slide have the same array arrangement rule. The size of the dot matrix area is 3.75mm×2mm, the dot pitch in the dot matrix is ​​250μm, the matrix: ...

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Abstract

The invention provides a gene chip and a kit for detecting invasive fungal pathogenic bacteria. The gene chip comprises a solid phase vector and an oligonucleotide probe fixed on the solid phase vector, wherein the probe contains DNA (Deoxyribose Nucleic Acid) sequences selected from 18S to 5.8S or 5.8S to 28S inside transcription regions (ITS) of rDNA and 5.8S rDNA of candida albicans, candida parapsilosis, candida tropicalis, candida kefyr, candida drusei, candida lusitaniae, candida glabrata, candida guilliermondii and cryptococcus neoformans as well as 5.8S rDNA of the nine fungus. The gene chip and the kit of the invention are utilized to detect nine kinds of common pathogenic bacteria, and have the advantages of simple and convenient operation, high accuracy and strong repeatability.

Description

technical field [0001] The invention relates to a gene chip and a detection kit for detecting nine common invasive pathogenic fungi. Background technique [0002] In recent years, due to the wide application of antibiotics, hormones, immunosuppressants and tumor radiotherapy and chemotherapy, the extensive use of various indwelling tubes and artificial valves in the body, and the increase of AIDS, the incidence of invasive fungal infection has increased significantly. and has become one of the important causes of death in critically ill patients. In the United States, the rate of deep fungal infection in hospitalized patients in 1990 was 1.9 times that of 1980, ranking first in the rate of infection by various pathogens. In 2004, it was reported that the rate of invasive fungal infection in hospitalized patients was 2.4 times that of the 1990s, and the mortality rate 29%; in Europe, between 1983 and 1997, invasive fungal infection accounted for 5.1% of all deaths, and betwe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C40B40/06C12Q1/68C12R1/74C12R1/725C12R1/72C12R1/645
Inventor 王磊喻群芳杨磊曹勃阳冯露
Owner TIANJIN BIOCHIP TECH CO LTD
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