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Fluorescent immunochromatographic test paper for detecting procalcitonin, and preparation method thereof

A technique of fluorescence immunochromatography and procalcitonin, which is applied in the field of biotechnology applications, can solve the problems of high cost, and achieve the effects of simple operation, high detection sensitivity and high efficiency

Pending Publication Date: 2020-02-11
南京欧凯生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, immunofluorescence requires an expensive immunofluorescence detector

Method used

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  • Fluorescent immunochromatographic test paper for detecting procalcitonin, and preparation method thereof
  • Fluorescent immunochromatographic test paper for detecting procalcitonin, and preparation method thereof

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preparation example Construction

[0037] The present invention also provides a method for preparing a fluorescent immunochromatographic test paper for detecting procalcitonin, comprising the following steps:

[0038] (1) Pretreatment of time-resolved immunofluorescence microspheres

[0039] After the time-resolved immunofluorescent microspheres were ultrasonically dispersed for 5 minutes, 50ul was taken, and the supernatant was removed after high-speed centrifugation at 14000rpm for 15min. The precipitate was washed with 190ul10mmol / L, pH6.0 MES buffer solution; Add 5ul of diimide and succinimide each, react at room temperature for 30min, centrifuge at 14000rpm for 15min at high speed, wash the precipitate with MES solution with a pH of 6.0 and resuspend to 500ul to obtain a time-resolved immunofluorescent microsphere solution;

[0040] (2) Preparation of time-resolved immunofluorescence microspheres combined with PCT monoclonal antibody

[0041] Add 50ug of PCT monoclonal antibody to the time-resolved immuno...

Embodiment 2

[0048] Example 2: Detection of PCT concentration by time-resolved fluorescence immunochromatography

[0049] Add different concentrations of human PCT antigen calibrator (100.0ng / ml, 50.0ng / ml, 20.0ng / ml, 10.0ng / ml, 5.0ng / ml, 1.0ng / ml in the test strip prepared by Example 1) , 0.5ng / ml, 0.2ng / ml, 0.05ng / ml, 0.02ng / ml, a total of ten concentrations, each concentration set three replicates, all of which are prepared by diluting 1.0mg / ml human PCT antigen with sample buffer) , after chromatography for 15min, read the fluorescence intensity ratio of the detection line and the control line (detection line detection value / control line detection value) ).

[0050] Take the concentration of PCT calibrator as the X-axis, and take the fluorescence intensity of the sample detection line as the Y-axis to establish an equation and fit it into a standard curve, such as figure 2 As shown, the R value is 0.9945.

[0051] The process specifically includes the following steps:

[0052] (1)...

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Abstract

The invention discloses fluorescent immunochromatographic test paper for detecting procalcitonin. The fluorescent immunochromatographic test paper comprises a PVC bottom plate, wherein a sample pad, acombination pad, a nitrocellulose membrane and a water absorption pad are sequentially arranged on the PVC bottom plate from left to right; one end of the sample pad is fixed on the PVC bottom plate,the other end of the sample pad is lapped on the combination pad, and a sample dripping hole is formed in the center of the sample pad; one end of the combination pad is fixed on the PVC bottom plate, and the other end of the combination pad is lapped on the nitrocellulose membrane; a detection line and a quality control line are sequentially arranged on the nitrocellulose membrane from left to right; and the left end of the water absorption pad is lapped on the nitrocellulose membrane. The fluorescent immunochromatographic test paper labels antibody proteins by means of time-resolved immunofluorescent microspheres, and carries out capture by using polyclonal antibodies, so that the detection sensitivity is higher; and the fluorescent immunochromatographic test paper greatly shortens thedetection time, is high in efficiency, and has good stability and repeatability.

Description

technical field [0001] The invention belongs to the application field of biotechnology, and relates to a fluorescent immunochromatographic test paper for detecting procalcitonin and a preparation method thereof. Background technique [0002] Procalcitonin (PCT) is a calcitonin precursor substance without hormone activity, which is composed of 116 amino acids and a glycoprotein with a molecular weight of 13KD. PCT is secreted by neuronal cells (including thyroid, lung and pancreatic tissue cells) and expressed, and is decomposed into calcitonin (CT)), carboxy-terminal peptide and amino-terminal peptide by enzymatic digestion. The half-life of PCT in the human body is 25-30h, with good stability, and its content in normal human serum is extremely low. In many cases leading to systemic inflammatory response syndrome (SIRS), such as bacterial infection, pancreatitis, burnt death, multiple injuries, the content of PCT in the blood is abnormally increased, while CT has no obvious...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/543G01N33/74G01N33/577
CPCG01N33/558G01N33/54313G01N33/74G01N33/577G01N2333/585
Inventor 戴瞻
Owner 南京欧凯生物科技有限公司
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