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Time-resolved fluorescence detection kit for monitoring 5-fluorouracil plasma-drug concentration in real time

A time-resolved fluorescence and detection kit technology, applied in the fields of medical biotechnology and molecular diagnosis, can solve the problems of low sensitivity, long detection time, complicated and cumbersome operations, etc., and achieves no radioactive pollution, wide detection range and good repeatability. Effect

Inactive Publication Date: 2017-09-05
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chromatography-mass spectrometry requires a large amount of serum samples, and the samples must be pre-treated before testing. The operation is complicated and cumbersome, and the detection time is long. It does not meet the requirements of real-time monitoring and is difficult to promote and apply.
Ultraviolet spectrophotometry also has a large demand for samples, low sensitivity, poor anti-interference ability, and poor specificity. It has little clinical significance and has been gradually eliminated.

Method used

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  • Time-resolved fluorescence detection kit for monitoring 5-fluorouracil plasma-drug concentration in real time
  • Time-resolved fluorescence detection kit for monitoring 5-fluorouracil plasma-drug concentration in real time
  • Time-resolved fluorescence detection kit for monitoring 5-fluorouracil plasma-drug concentration in real time

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] A time-resolved fluorescence detection kit for real-time monitoring of blood concentration of 5-fluorouracil, including calibrator 5-FU with a concentration of 0, 10, 50, 100, 750 and 1500ng / mL, and the composition is 2mg / ml BSA, 0.1% NaN 3 , 50mM pH 7.8 Tris-HCl buffer calibrator dilution; europium-labeled 5-FUA-OVA antigen; anti-5-FU mouse polyclonal antibody; components are 50Mm pH 7.8 Tris-HCl, 1.5% PEG6000, 0.2% BSA, 0.1% Proclin300, 0.02% bovine IgG, 0.1% Tween 20 and 0.84% ​​NaCl assay buffer; composition is 1.25mmol / L pH 7.8 Tris-HCl, 2.5% Tween 20 and 21% NaCl 25× washing solution; composition is 15umol / L β-naphthoyl trifluoroacetone, 50umol / L trioctylphosphine oxide, 0.1% TritonX-100, 0.6% acetic acid, potassium hydrogen phthalate to adjust pH3.0~3.2 enhancement solution; goat anti-mouse IgG-coated reaction plate; also includes 3 coverslips and 1 copy of instructions.

[0049] Wherein, the preparation steps of europium-labeled 5-FUA-OVA antigen are as follow...

Embodiment 2

[0077]The methodological verification of the test kit of the present invention, according to the conventional manufacturing and verification procedures in the art, the test kit prepared above is tested, and the results are as follows:

[0078] (1) Analytical sensitivity and linear range: measure 10 times with the zero reference standard as the sample, and calculate its fluorescence value and standard deviation. The concentration obtained by substituting the fluorescence value obtained by subtracting 2 times the standard deviation from the mean value of the fluorescence measurement at this point into the standard equation is its analytical sensitivity. The analytical sensitivity of this reagent is determined to be 2.1ng / ml. The antigen was diluted into different concentrations for determination, and the linear range of the measured standard curve was 2.1-1500ng / ml, the standard curve was LOG(Y)=5.024-0.194LOG(X), R=0.9986, see the standard curve figure 2 .

[0079] (2) Precis...

Embodiment 3

[0086] Test kit of the present invention and liquid chromatography-mass chromatography (LC-MS / MS) clinical blood sample measurement value comparison: use test kit of the present invention and liquid chromatography (LC-MS / MS) to simultaneously test 110 parts of serum Samples are tested. Measure the 5-FU concentration result of the blood sample by the method of the present invention as the abscissa, and use the 5-FU concentration result measured by the LC-MS / MS method as the ordinate to do regression analysis, and the correlation equation is: Y=0.981X-3.267, the correlation coefficient r=0.981 ( image 3 ). The results of statistical processing show that the method of the present invention has a significant correlation with the measured value of the clinical blood sample by the LC-MS / MS method.

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Abstract

The invention discloses a time-resolved fluorescence detection kit for monitoring 5-fluorouracil plasma-drug concentration in real time. The kit comprises calibrator 5-FU, calibrator dilution liquid, europium-standard 5-FUA-OVA antigens, anti-5FU murine polyclonal antibodies, analysis buffer solution, scrubbing solution, enhancement solution and a goat-anti-mouse IgG (immune globulin G) package reaction plate. The kit can quantitatively detect plasma-drug concentration of the 5-fluorouracil in peripheral blood of chemotherapy patients, the testing results show that the kit has the advantages that the kit is high in sensitivity, good in stability and reliability, high in efficiency, simple and convenient to operate, wide in detection range and free from radioactive pollution, time is saved and the like, the sensitivity is 2.1ng / ml, chemotherapy drugs are timely monitored, the kit is suitable for plasma-drug monitoring of clinical fluorouracil drugs, and clinical medication is guided.

Description

technical field [0001] The invention relates to the fields of medical biotechnology and molecular diagnosis, in particular to a time-resolved fluorescence detection kit for real-time monitoring of blood drug concentration of 5-fluorouracil. Background technique [0002] 5-fluorouracil (5-fluorouracil, 5-FU) is a broad-spectrum, highly effective drug of choice for the treatment of gastrointestinal and respiratory tumors, and 5-FU-based combination therapy is also the standard adjuvant treatment for colorectal cancer after surgery . Although 5-FU has a significant curative effect, it has side effects such as myelosuppression and mucositis, which limit its clinical application. Clinical pharmacoepidemiological surveys show that there are obvious individual differences in its toxic and side effects, which is related to the different individual expression levels of the catabolic rate-limiting enzyme-dihydropyrimidine dehydrogenase (Dihydropyrimidinedehydrogenase, DPD). The toxi...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N21/64
CPCG01N33/533G01N21/6408G01N21/643G01N2021/6439
Inventor 吴英松刘天才林冠峰梁君瑜
Owner SOUTHERN MEDICAL UNIVERSITY
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