Detection method for human tissue kallikrein in body fluid and its use

A technology of kallikrein and assay method, applied in the field of diagnosis of cerebral ischemia, can solve the problems of chronic ischemia, not obvious surgical indications, unsuitable for general screening of risk groups of stroke, etc., and achieve low detection cost Effect

Inactive Publication Date: 2004-09-29
赵立
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Problems solved by technology

[0004] Epidemiological studies have found that patients with high carotid artery stenosis are prone to stroke, and surgical recanalization of obstructed arteries can reduce the risk of stroke; however, for patients with low to moderate carotid artery stenosis, comprehensive consideration of perioperative risk and artery The benefits of recanalization, the surgical indications are not obvious; and patients with carotid artery stenosis may also develop acute or chronic ischemia, resulting in progressive brain function damage
Cerebral perfusion cannot be accurately estimated from cerebral angiography
Even though there are currently more accurate diagnostic techniques than cerebral angiography, this technique is often expensive and not suitable for universal screening of stroke risk populations

Method used

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  • Detection method for human tissue kallikrein in body fluid and its use
  • Detection method for human tissue kallikrein in body fluid and its use
  • Detection method for human tissue kallikrein in body fluid and its use

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Embodiment 1

[0033] Purification of tissue kallikrein from human urine

[0034]Tissue kallikrein can be extracted from human urine by affinity chromatography (Shimamoto K., Chao, J., Margolius H.S. (1980). J. Clin. Endocinology and Metabolism, 51:840-848). Collect 12 liters of urine and concentrate it to 300 ml by ultrafiltration, and store it at -20 degrees. Repeated freezing and thawing, filtered through three layers of gauze and Nitex membrane, centrifuged at 5000g for 30 minutes, the precipitate was salted out with 35-80% ammonium sulfate, collected by centrifugation, redissolved and dialyzed, the dialysate passed through DE-52 cellulose chromatography column, Elute with 0.1-0.5M sodium chloride-0.1M phosphate buffer (pH 6.5), collect the elution peak with Tos-Arg-Ome esterase activity, for 0.2M sodium chloride-0.1M phosphate buffer (pH6.5) for dialysis, the dialysate is passed through the agarose gel affinity column with aprotinin as the ligand, eluted with the dialysis buffer until ...

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Abstract

The present invention relates to the diagnosis technology of cerebral ischemia, and is especially the detection method of human tissue kallikreinase in body fluid and its application in cerebral ischemia diagnosing kit. The detection method includes immunizing globin specifically with human tissue kallikreinase or human tissue kallikreinase segment for reaction with the tissue kallikreinase in biological sample to form immune compound; detecting the immune compound formed in the biological sample and comparing with reference sample to obtain the content of tissue kallikreinase in biological sample. One new type of biological cerebral ischemia marker with low detection cost and fast and simple screening is revealed. The cerebral ischemia diagnosing kit with specifically immunized human tissue kallikreinase or human tissue kallikreinase segment is prepared therewith, and may be used in fast estimation of the danger / benefit ratio of operating dredging of carotid, etc.

Description

technical field [0001] The invention relates to a diagnostic technique for cerebral ischemia, in particular to a method for measuring human tissue kallikrein in body fluid and its application in a diagnostic kit for cerebral ischemia. Background technique [0002] Tissue kallikrein is a protease found in blood, other body fluids, and tissues. Its active metabolite is kinin, which has strong vasodilatory activity; tissue kallikrein can exist in free form, or It exists in a complex with its natural inhibitors, such as tissue kallikrein inhibitors. The tissue kallikrein-kinin system in brain tissue is involved in the central regulation of cardiovascular function (Madeddu P. et al., Hypertension 1996; 28: 980-989; Emanueli C. et al., Br. J. Pharmacol. 1999; 126: 1769-1776), the product of tissue kallikrein, kinin, dilates the cerebral vascular network (Toda N., Am. J. Physiol., 1997; 222: H267-H274). The tissue kallikrein-kinin system in brain tissue can be activated by tissue...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573
Inventor 伊曼纽李·康斯坦察唛德杜·保罗赵朱丽赵立宝库·保罗
Owner 赵立
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