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Process for testing gene chip used to detect bacteremia

A technology of gene chip and inspection method, which is applied in the field of target gene DNA sequence amplification and identification, can solve the problems of poor accuracy and missing pathogenic bacteria, and achieve the effect of ensuring accuracy and increasing the positive rate

Inactive Publication Date: 2006-06-28
NANKAI UNIV +1
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  • Description
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Problems solved by technology

However, fewer probes are included, only 22 types of pathogenic bacteria are covered, many pathogenic bacteria will be missed, and the accuracy is poor

Method used

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  • Process for testing gene chip used to detect bacteremia
  • Process for testing gene chip used to detect bacteremia
  • Process for testing gene chip used to detect bacteremia

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Embodiment

[0031] Collect blood samples, 0.1-0.5ml. Take 0.1ml of whole blood and place it in an eppendorf centrifuge tube, add 1ml of sterile water and let stand at room temperature for 5 minutes, then centrifuge at room temperature: 12000rpm for 5 minutes, discard 900μl supernatant, add 400μl lysis buffer (lysis Buffer: 10mM Tris.HCL pH8.0, 10mM EDTA, 0.5% SDS) after mixing, add 6μl of 20mg / ml proteinase K (Sigma or Shanghai Shenggong Company), mix evenly, put it in a water bath at 56°C for 30 minutes, and then Add an equal volume of TE solution saturated with phenol reagent, shake vigorously, centrifuge at 8000r / min room temperature for 3 minutes, absorb the supernatant, repeat the phenol extraction once, absorb the supernatant, add 1 / 10 volume of sodium methanolate (3mol / l ), mix, add an equal volume of ice-cold isopropanol, after mixing, centrifuge: 12,000rpm, room temperature, 5 minutes, pour off the supernatant, add 70% cold ethanol to shake and wash once, centrifuge: 12,000rpm, r...

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Abstract

A process for detecting bacteremia with gene chip includes such steps as separating the DNA of pathogenic bacteria in blood, PCR amplification, labelling target DNA, hybridizing with the probe array on gene chip and enzyme-linked immunosorbant assay to realize the classification and appraised of pathogenic bacteria. Its advantages are high speed and high correctness.

Description

technical field [0001] The present invention relates to the detection method of the gene chip for bacteremia detection, especially the technology for the separation of pathogenic bacteria DNA in blood, the amplification and identification of target gene DNA sequence, which can be extended to human body, livestock and other products based on the gene chip. Organ infection germ detection, environment, sanitation detection and customs quarantine technology. Background technique [0002] Normal human blood is sterile. When bacteria invade the blood, it can cause bacteremia and even sepsis in humans. This disease is a serious and critical systemic infection, and bacteriological examination is of great significance to the prevention and treatment of the disease. There are two main traditional identification methods for pathogenic bacteria in blood. One is the most traditional culture, separation, and identification system. Venous blood is collected aseptically, and immediately in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 黄熙泰侯亭
Owner NANKAI UNIV
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