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Homogeneous phase multi-index fluorescence/chemiluminescence measuring method and application thereof

A chemiluminescence and fluorescence measurement technology, applied in the field of blood testing, can solve the problems of single detection marker, high diagnostic cost, long experimental period, etc., and achieve the effects of simple operation, low diagnostic cost, and fast detection time.

Inactive Publication Date: 2012-12-12
FUDAN UNIV
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Problems solved by technology

[0005] The purpose of the present invention is to provide a homogeneous multi-index fluorescence / chemiluminescence assay method in order to overcome the shortcomings of the prior art, such as single detection markers, long experimental period, large amount of serum, and high diagnostic costs, especially related to a tumor and other major Fluorescence / chemiluminescence simultaneous detection technology for multiple serum markers of diseases

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  • Homogeneous phase multi-index fluorescence/chemiluminescence measuring method and application thereof

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Embodiment 1

[0033] 1) Binding the coated antibody to polystyrene microspheres to prepare the carrier-antibody complex

[0034] First, the carboxypolystyrene microspheres were transferred into a 1.5 mL centrifuge tube, washed three times by centrifugation at 12,000 rpm with an imidazole solution (0.1 M) of pH=6.0, and then activated by adding 100 μL of imidazole solution containing 1 mg EDC at 37° C. for 30 min. Then the activated polystyrene microspheres were washed three times by centrifugation with imidazole solution, and finally dispersed in 100 μL imidazole solution. Then add CEA, NSE and Cyfra21-1 coating antibody, and react at 37°C for 1h. After washing three times with PBSTW washing solution, 200 μL of PBS solution containing 2% BSA was added, and blocked at 37° C. for 1 h. After the polystyrene microsphere-antibody complexes were washed, they were stored in 200 μL of PBS solution containing 2% BSA for later use.

[0035] 2) Preparation of digoxin anti-CEA labeled antibody

[00...

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Abstract

The invention belongs to the field of blood testing, and relates to a homogeneous phase multi-index fluorescence / chemiluminescence measuring method and application thereof. The method comprises the steps of: fixing a plurality of specificity recognition monoclonal antibodies on a polystyrene microsphere, and reacting with a plurality of labeled antibody cores modified by haptens after combining with carcinoembryonic antigens, cell keratin fragments 19, neuron specificity enolase and other multiple markers; reacting with horse radish peroxidase modified by corresponding hapten antibodies, alkaline phosphatase and quantum dots, and measuring a plurality of markers on the same instrument by combining with detections of chemiluminescence, fluorescence and the like. The method has the advantages of small blood serum consumption, rapid detection time, low diagnosis cost and simple operation, is suitable for detecting a plurality of popularized indexes simultaneously, and can provide auxiliary judgment basis for the early diagnosis of critical illnesses of tumor and the like.

Description

technical field [0001] The invention belongs to the field of blood testing, and relates to a new technology for simultaneous detection of fluorescence / chemiluminescence, in particular to a homogeneous multi-index fluorescence / chemiluminescence measurement method and its application. In particular, it relates to a fluorescence / chemiluminescence simultaneous detection technology for multiple serum markers of major diseases such as tumors. Background technique [0002] Epidemiological studies have shown that not only is the cancer mortality rate in my country showing an obvious upward trend, but it also has the characteristics of high incidence in developing countries and developed countries. There are four main reasons: (1) Population aging. Fifty years ago, the average life expectancy in my country was about 49 years old, but now it is more than 70 years old, which is very close to the level of developed countries. Aging leads to a natural increase in senile diseases; (2) Ind...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G01N21/76G01N33/574G01N33/577G01N33/533
Inventor 李欢曹志娟刘彩云卢建忠
Owner FUDAN UNIV
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