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Application of Mfsd2a (major facilitator superfamily domain-containing protein 2a) to preparation of product for diagnosing purulent meningitis

A suppurative meningitis and product technology, applied in measuring devices, individual particle analysis, scientific instruments, etc., can solve problems such as cerebrospinal fluid limitation, difficulty in diagnosing suppurative meningitis, low birth weight, etc.

Inactive Publication Date: 2017-12-01
QILU CHILDRENS HOSPITAL OF SHANDONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The diagnosis of purulent meningitis mainly depends on the examination of cell count in cerebrospinal fluid, but it is sometimes quite difficult to confirm the diagnosis of purulent meningitis in clinical work, because the early clinical symptoms are sometimes mild (especially for infants who cannot describe their symptoms) , access to cerebrospinal fluid through invasive procedures such as lumbar puncture is clinically limited, especially in newborns born prematurely, with fat or low birth weight

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  • Application of Mfsd2a (major facilitator superfamily domain-containing protein 2a) to preparation of product for diagnosing purulent meningitis
  • Application of Mfsd2a (major facilitator superfamily domain-containing protein 2a) to preparation of product for diagnosing purulent meningitis
  • Application of Mfsd2a (major facilitator superfamily domain-containing protein 2a) to preparation of product for diagnosing purulent meningitis

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Embodiment 1

[0016] In vitro, Escherichia coli, the main pathogen of bacterial meningitis in children, was used to stimulate human brain microvascular endothelial cells (HBMECs) in vitro, and it was confirmed that Escherichia coli stimulated HBMECs in vitro (0.5916 ± 0.041 Compared with the control group (0.278±0.043 ), the expression of Mfsd2a was significantly increased (P=0.0060) ( figure 1 ).

[0017] In vivo experiments, Escherichia coli was injected into the animal model of meningitis in newborn C57BL / 6J mice, and the changes of Mfsd2a content in the blood and cerebrospinal fluid of the mice were detected, confirming that Escherichia coli stimulated newborn C57BL / 6J mice ( 1.824±0.323 Int / mm 2 ) Mfsd2a content in peripheral blood was higher than that of the control group (0.314±0.141 Int / mm 2 ) was significantly higher than that of Mfsd2a (P=0.0055). At the same time, newborn C57BL / 6J mice stimulated by Escherichia coli (2567.0±434.8 Int / mm 2 ) Mfsd2a content in cerebrospinal flu...

Embodiment 2

[0021] Further preferably, in the human cerebrospinal fluid of newborns infected with Escherichia coli, the change of Mfsd2a content in the human cerebrospinal fluid is detected, and the Mfsd2a content in the human cerebrospinal fluid of confirmed infection of Escherichia coli (12650 ± 2734 Int / mm 2 ) and the control group (3953±424 Int / mm 2 ) was significantly higher than that of Mfsd2a (P=0.0348)) (Figure 4).

[0022] Western blot method was used to detect the expression level of Mfsd2a protein in HBMECs, mouse brain and CSF and clinical cerebrospinal fluid samples. The brain tissue was suspended in a buffer containing 1% protease inhibitor (Sigma-Aldrich) lysate (Cell Signaling Technology Company), and then centrifuged at 12,000 g for 15 minutes at 4°C. All protein products were stored at -80°C. 30µL mouse CSF was diluted with 400µL PBS for protein electrophoresis. Clinical cerebrospinal fluid samples also need to be centrifuged at 12,000 g for 15 minutes at 4°C and stor...

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Abstract

The invention provides application of Mfsd2a (major facilitator superfamily domain-containing protein 2a) to preparation of a product for diagnosing purulent meningitis. On one hand, the invention provides a biological product; the product can be used for detecting the concentration change of the Mfsd2a; on the other hand, the invention provides a method for labeling by adopting fluorescence; the Mfsd2a is used as a detection marker, and a circulating cerebrovascular endothelial cell which is separated from human peripheral blood by adopting a UEA I (Ulex Europaeus Agglutinin I) magnetic bead method is used for experimenting to diagnose the purulent meningitis in vitro. The invention provides a novel molecule Mfsd2a for clinically diagnosing the purulent meningitis and develops a method for diagnosing the purulent meningitis in vitro based on a peripheral blood specimen by taking the Mfsd2a as the marker.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis, and in particular relates to the application of Mfsd2a in preparing products for diagnosing suppurative meningitis. Background technique [0002] Suppurative meningitis is an infectious disease of the central nervous system caused by purulent bacteria, mainly meningitis. diplococci etc. Suppurative meningitis causes pathological changes such as blood-brain barrier dysfunction and neutrophil infiltration. Neonatal encephalitis is one of the three most common neonatal diseases, causing approximately 393,000 neonatal deaths per year worldwide. [0003] The diagnosis of purulent meningitis mainly depends on the examination of cell count in cerebrospinal fluid, but it is sometimes quite difficult to confirm the diagnosis of purulent meningitis in clinical work, because the early clinical symptoms are sometimes mild (especially for infants who cannot describe their symptoms) However, acc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/10
CPCG01N15/10G01N2015/1006
Inventor 王世富张乐海王素兰李政于风岭
Owner QILU CHILDRENS HOSPITAL OF SHANDONG UNIV
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