GFAP detection method based on fluorescence resonance energy transfer method

A technology of energy resonance transfer and detection method, which is applied in fluorescence/phosphorescence, biological testing, measuring devices, etc., can solve the problems of long inspection time and cumbersome steps, and achieve the effects of shortened detection time, high specificity, and good accuracy

Active Publication Date: 2017-04-19
SHAANXI MYBIOTECH CO LTD
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Problems solved by technology

[0004] At present, the detection of GFAP in clinical diagnosis is mainly the ELISA experimental method, which has been widely used in the past two decades because of its specificity and high sensitivity, but the method is cumbersome and takes a long time to test

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  • GFAP detection method based on fluorescence resonance energy transfer method
  • GFAP detection method based on fluorescence resonance energy transfer method
  • GFAP detection method based on fluorescence resonance energy transfer method

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Embodiment Construction

[0022] The present invention will be further described in detail below in conjunction with specific embodiments, which are explanations of the present invention rather than limitations.

[0023] The disclosed GFAP detection method based on the fluorescence energy resonance transfer method of the present invention requires simultaneous recognition by three antibodies, has higher specificity than Elisa, and avoids some false positives of the Elisa method. At the same time, direct detection by FRET between two fluorescent-labeled antibodies saves the steps of multiple washings and substrate color development in Elisa, greatly shortening the detection time.

[0024] The GFAP detection method based on fluorescence energy resonance transfer method disclosed by the present invention comprises the following operations:

[0025] 1. Collect serum:

[0026] Whole blood samples were left at room temperature for 2 hours or overnight at 4°C and then centrifuged at 1000×g for 20 minutes, an...

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Abstract

The present invention discloses a GFAP detection method based on a fluorescence resonance energy transfer method, wherein a micro-pore plate is coated with a purified glial fibrillary acidic protein (GFAP) antibody to prepare a solid-phase antibody, glial fibrillary acidic protein (GFAP) is sequentially added to the monoclonal antibody-coated micro-pores, combination with Alexa Fluor488 labeled and Alexa Fluor594 labeled GFAP antibody to form an antibody-antigen two-fluorescent-labeling-antibody complex, absorbance (OD value) is determined by using a fluorescent enzyme analyzer according to the FRET principle, and the human glial fibrillary acidic protein (GFAP) content in the sample is calculated by using a standard curve. According to the present invention, with the use of the three kinds of the antibodies to simultaneously identify, such that the specificity is high, the accuracy is good, the detection time is substantially shortened compared to the existing method, the reference information is provided for the clinical diagnosis, the clinicians can be helped to diagnose and assess the progression of stroke, and the prognosis of patients can be improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a GFAP detection method based on fluorescence energy resonance transfer (FRET). Background technique [0002] Epidemiological surveys show that cerebrovascular disease, heart disease, and malignant tumors constitute the three major causes of human death. Compared with developed countries in the West, the morbidity and mortality of cerebrovascular diseases in my country are significantly higher than those of cardiovascular diseases. It is estimated that there are about 2 million new stroke patients in China every year; about 1.5 million patients die of stroke every year; 6 million to 7 million surviving patients; and 70% to 80% of the survivors are left with paralysis and aphasia. Severe disabilities, such as disability, bring heavy burdens to society and families. Among them, cerebral hemorrhage is a common and frequently-occurring disease with high mortality and disabi...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/68
CPCG01N21/6486G01N33/68
Inventor 闫亚平秦李娜李科
Owner SHAANXI MYBIOTECH CO LTD
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