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

Inactive Publication Date: 2018-11-06
刘健刚
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0009] The purpose of the present invention is to solve the above technical problems, and to provide a method for rapid detection of postoperative intracranial bacterial infection by multiple real-time fluorescent PCR. The corresponding bacterial species are detected, the specificity and sensitivity of the detection method are high,

Method used

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  • Method for rapid detection of postoperative intracranial bacterial infection by multiple real-time fluorescent PCR (Polymerase Chain Reaction)
  • Method for rapid detection of postoperative intracranial bacterial infection by multiple real-time fluorescent PCR (Polymerase Chain Reaction)
  • Method for rapid detection of postoperative intracranial bacterial infection by multiple real-time fluorescent PCR (Polymerase Chain Reaction)

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Experimental program
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Embodiment

[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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Abstract

The invention provides a method for rapid detection of postoperative intracranial bacterial infection by multiple real-time fluorescent PCR (Polymerase Chain Reaction) and belongs to the technical field of molecular biology. The method adopts a multiple fluorescent PCR technology to detect the cerebrospinal fluid of a patient with intracranial surgery fever so as to realize the typing detection ofa plurality of common intracranial infection bacteria. The method provided by the invention can realize rapid detection of postoperative intracranial bacterial infection, can quickly identify commonbacterial types in patients with intracranial infection after surgery, and solves the problems that a culture method can not identify new strains and can not culture bacteria. The detection method hashigh specificity and sensitivity, reduces the false positive of PCR detection, shortens the detection period, can realize timely and rapid diagnosis of intracranial bacterial infection, and has significant significance for blocking clinical infection and guiding medication.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and medicine, and relates to a fluorescence detection method, in particular to a multiple real-time fluorescence PCR rapid detection method for postoperative intracranial bacterial infection. Background technique [0002] Intracranial infection is one of the common serious complications after neurosurgical craniotomy, which can cause damage to nervous system function, and its incidence rate is about 2.6% to 30%. Once it occurs, it not only seriously affects the prognosis of patients, but also the symptoms are difficult to control and often life-threatening. Because intracranial bacterial infection often affects the brain, spinal cord and adjacent anatomical tissues, patients are often accompanied by symptoms such as high intracranial pressure, disturbance of consciousness, cerebral edema, and epilepsy. A serious burden has been imposed on their families and society. And if these patie...

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Application Information

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IPC IPC(8): C12Q1/6851C12Q1/14C12Q1/10C12R1/445C12R1/01C12R1/46C12R1/19C12R1/38C12R1/22
CPCC12Q1/6851C12Q2531/113C12Q2537/143C12Q2561/113C12Q2563/107
Inventor 刘健刚
Owner 刘健刚
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