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Method for identifying 28 frequent phthogenic bacteria for clinical bacteremia

A technology for pathogenic bacteria and bacteremia, applied in the fields of molecular biology and biochemistry, can solve the problems of high price, many influencing factors and high price, and achieve the effect of shortening detection time, improving identification efficiency and speed.

Inactive Publication Date: 2006-08-09
ZHEJIANG UNIV
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AI Technical Summary

Problems solved by technology

However, the traditional solid-phase biochip has gradually exposed its shortcomings in the long-term use process: (1) poor flexibility: biochips are prefabricated according to a certain pattern, and cannot adapt to the ever-changing needs of patients; (2) ) detection takes a long time: because the reaction is carried out on glass or membrane substrates, it must be washed repeatedly to remove excess reaction starters, intermediate products and hybridization buffers, etc.; (3) repeatability Not ideal: Hybridization on glass or membrane substrates has many influencing factors, and it is difficult to strictly control the experimental conditions
(4) Expensive: The biochips currently in clinical use are all expensive, which is unbearable for patients

Method used

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Examples

Experimental program
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Embodiment Construction

[0030]1. Collection of strains: Most of the 91 purely cultured strains used were clinical isolates and were collected from the Affiliated Hospital of Zhejiang University from April 2004 to August 2005. Including 6 standard strains, namely Klebsiella pneumoniae (ATCC700603), Escherichia coli (ATCC25922), Enterococcus faecalis (ATCC29212), Pseudomonas aeruginosa (ATCC27853), Staphylococcus aureus (ATCC25923), monocyte Listeria monocytogenes (NICP BP54001). ATCC refers to the American Culture Collection; NICPBP refers to the National Institute for the Control of Pharmaceutical and Biological Products.

[0031] 2. Obtain strain DNA

[0032] Pick a single colony and put it in 50 μl extraction lysate (10mM Tris-HCl pH value 7.6, 5mM EDTA, 0.5% SDS, 0.22μm membrane filtration treatment), boil at 100°C for 10min, centrifuge at 12000r / min for 5min, take 2μl supernatant as PCR template.

[0033] 3. 23S rDNA amplification and labeling of strains to be tested

[0034] The P6 / P10r prim...

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Abstract

This invention relates to a method for identifying 28 kinds of common pathogens of clinical bacteremias, which crosslinks 30 types of specific probes against the 28 kinds of common pathogens on 30 kinds of different fluorescent microspheres to be reacted with the being tested specimens then to be reacted with the report molecules labeled by fluorescent elements to test the specific 23SrDNA of them on a fluorescent test device.

Description

technical field [0001] The invention belongs to the field of molecular biology and biochemistry of biotechnology, relates to a suspension chip technology for identifying common pathogenic bacteria in clinical bacteremia, is suitable for identification of bacterial strains isolated clinically, and accurately and reasonably conducts antibacterial treatment for patients with clinical bacteremia Drug therapy provides guidance. Background technique [0002] Isolation of bacteria from blood cultures indicates a serious infection requiring immediate antimicrobial therapy. Different pathogenic bacteria have different sensitivities to drug types and concentrations, and the key to successful treatment lies in the early application of appropriate and sufficient drugs. It usually takes 1 to 3 days to isolate and identify bacteria in blood culture, during which improper medication often makes the patient's condition worse or increases the drug resistance of the strain. Especially in th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 陈智侯晓丽朱海红
Owner ZHEJIANG UNIV
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