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.
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[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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