Nucleic acid sequence, method and kit for diagnosis of meningitis pathogenic bacteria
A technology for pathogenic bacteria nucleic acid and meningitis, which is applied in biochemical equipment and methods, microbial determination/inspection, sugar derivatives, etc. higher question
Inactive Publication Date: 2005-12-21
天津市中敖畜牧业有限公司
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Problems solved by technology
However, it takes at least three days to more than a week to directly culture bacteria, and antibody identification is mostly one-to-one mode. Using this method to identify bacteria species requires high costs and is likely to cause waste of medical resources.
[0004] Based on the shortcomings of the current diagnosis technology for meningitis pathogens, it can be known that the rapid and accurate detection of meningitis pathogens cannot be achieved due to factors such as high cost and long detection and inspection time. Therefore, in order to improve medical standards and benefits, currently There is a great need for a high-efficiency method and kit for detecting meningitis pathogenic bacteria that can overcome the above-mentioned shortcomings
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[0144] The following examples serve to illustrate the invention. These examples are not intended to limit the scope of the invention in any way, but rather serve as an indication of how to practice the materials and methods of the invention. The best implementation state of the meningitis pathogenic bacteria diagnostic chip of the present invention comprises the following steps:
[0145] A. Extraction of DNA of pathogenic bacteria:
[0146] (1) A pathogen colony is screened with 500ul solution I, wherein solution I is:
[0147] 50mM Glucose
[0148] 10mM EDTA
[0149] 25mM Tris-HCl pH=8.0
[0150] the mixed solution.
[0151] (2) For G(+) pathogenic bacteria, add 1 mg / ml lysozyme and incubate at 37°C for 30-60 minutes; for G(-) pathogenic bacteria, skip directly to step (3).
[0152] (3) Add 50 μl of 10% SDS and incubate at 65° C. for 30-60 minutes.
[0153] (4) Add RNase 4mg / mL and incubate at 37°C for 30-60 minutes.
[0154] (5) Add 100 μl of 5M KAc and 300 μl of CHC...
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The invention provides a nucleic acid sequence, method and kit for diagnosing meningitis pathogens, which can quickly diagnose 20 kinds of meningitis pathogens. The principle is to compare the DNA sequences of various pathogens, select special fragments, and find out Corresponding common primers, and design specific probe fragments for various pathogenic bacteria, design 3 specific probe fragments for each pathogenic bacteria, and then place these probe points on the polymer to make meningitis application chip. The DNA of the pathogenic bacteria in the clinical specimen is extracted with a reagent, amplified by calibration, reacted with the meningitis pathogenic bacteria diagnostic chip of the present invention, and the infected fungus can be identified with the naked eye. Compared with the traditional method, the chip detection method of the present invention can quickly detect various infectious bacteria species, and the cost is saved by at least half.
Description
technical field [0001] The invention relates to a nucleic acid sequence, a method and a kit for diagnosing meningitis pathogenic bacteria, capable of targeting Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus saprophyticus and Streptococcus agalactiae ), Streptococcus.Pyogens, Streptococcus.Pneumoniae, Enterococcus faecium, Enterococcus faecalis, Mycobacterium tuberculosis, Pneumophilic Legionnaires Legionellapneumophilia, Listeria monocytogenes, Escherichia coli, Klebsiella pneumoniae, Serratia marcescens, Enterobacter cloacae cloacae), Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Proteus mirabilis, Haemophilus influenzae and Neisseriameningitidis for rapid diagnosis of 20 meningitis pathogens, the principle is to Compare the DNA sequences of various pathogenic bacteria, select special fragments, find out the corresponding common primers, and design specific probe fragments for various pathogenic bacteria, design 3 specific probe fragments for each p...
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IPC IPC(8): C07H21/00C12Q1/04C12Q1/68
Inventor 许博翔江雅铃黄献龙赵守宇
Owner 天津市中敖畜牧业有限公司