Method for rapid detection of postoperative intracranial bacterial infection by multiple real-time fluorescent PCR (Polymerase Chain Reaction)
A real-time fluorescence, bacterial infection technology, applied in the fields of molecular biology and medicine, can solve the problems of reducing PCR false positives, shortening the detection cycle, unable to cultivate bacteria, etc., achieving high specificity and sensitivity, shortening the detection cycle, and reducing false positives Effect
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[0050] 1 Materials and Instruments
[0051] 1.1 Materials
[0052] Tested strains: Staphylococcus aureus, Enterococcus faecalis, Streptococcus pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii, a total of 7 strains were controlled by the Shanghai Municipal Center for Disease Control and Prevention. Donated by the center, five clinical isolates of Mycobacterium tuberculosis, Enterobacter cloacae, Enterobacter aerogenes, herpes simplex virus and cytomegalovirus were provided by the Third Affiliated Hospital of Guangzhou Medicine.
[0053] All strains were inoculated in beef extract peptone medium, cultured at 37°C for 18 hours, and transferred for 3 times before use.
[0054] 1.2 Reagents and instruments
[0055] Reagents: Hot Start Taq DNA Polymerase, 10×Hot Start PCR buffer, dNTP, MgCl 2 , Shanghai Zhaowei Technology Development Co., Ltd.; EcoR I, DNase Ⅰ, Bao Biological Engineering (Dalian) Co., Ltd.; PCR primers were s...
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