Ultra-sensitive fluorescence quenching immunosensor of detecting human cTnI in serum and detection method

A non-fluorescent, ultra-high technology, applied in the field of immunochromatography, it can solve the problems of unstable quantitative detection results, limitations, high false positive rate, and achieve the effect of solving diffusion and blooming

Active Publication Date: 2020-01-03
GUILIN UNIVERSITY OF TECHNOLOGY
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Problems solved by technology

[0005] However, due to quality differences, immunochromatographic test strip products generally have the defects of high false positive rate, low sensitivity, high detection limit, and unstable quantitative test results. Sensitive quantitative chromatography test strips, but its application is mostly limited to the stage of small tests in the laboratory

Method used

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  • Ultra-sensitive fluorescence quenching immunosensor of detecting human cTnI in serum and detection method
  • Ultra-sensitive fluorescence quenching immunosensor of detecting human cTnI in serum and detection method
  • Ultra-sensitive fluorescence quenching immunosensor of detecting human cTnI in serum and detection method

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Embodiment 1

[0032] In this example, FITC is used as the fluorescent signal source, and the required two-phase buffer system is designed according to the charging properties of FITC. Based on this, the fluorescence quenching immunosensor is prepared and tested for different concentrations of human cTnI in serum. The test includes test strips Fluorescence emission spectrum characterization of test lines and quality control lines.

[0033] 1) Prepare 10mMmol / L PBST coating solution, specifically Tween-20 containing 0.5% final volume concentration, 5% saturated sucrose, 5% methanol, 2% BSA, and set the volume to a concentration of 10mM / L L, the PBS Buffer solution that pH value is 7.2;

[0034] 2) 1) the 10mMmol / L PBST coating solution with a final volume concentration of 90%, 10% of 5mol / L NH 4 Cl prepares FITC with two-phase buffer system;

[0035] 3) with 1) described 10mMmol / L PBST coating fluid as solvent, prepare respectively human cTnI antigen solution, mouse anti-human cTnI antibody...

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Abstract

The invention discloses an ultra-high-sensitivity fluorescence quenching immunosensor of using human troponins I (cTnI) in serum as target proteins and a detection method. The immunosensor is mainly characterized in that the established immunosensor realizes ultra-high-sensitivity detection on the human cTnI in the human serum. On the basis of a lateral-flow immunity chromatography platform, the fluorescence quenching immunosensor of using the human cTnI as model targets is established in the invention, a detection limit on human IgG is determined as 16.26 fg / mL through calculation, results can be obtained averagely after 6 minutes, and ultra-high-sensitivity POCT fast-immunoassay on the above-mentioned target antigens is realized. The lateral-flow immunity chromatography platform disclosed by the invention realizes preparation of lower cost and a shorter process on the premise of ensuring stability and ultra-high sensitivity.

Description

technical field [0001] The invention belongs to the field of immunochromatography, and in particular relates to an ultrahigh-sensitivity non-fluorescence-labeled immunoassay sensor which uses human troponin I (cTnI) in serum as a target protein. Background technique [0002] Acute Myocardial Infarction (AMI) is a sudden clinical disease. In addition to symptoms such as chest pain, nausea, autogenous fever, and unstable heart rhythm, the distribution concentration of corresponding myocardial infarction markers in the body There will also be a large change, which makes myocardial infarction markers a key indicator for the diagnosis of AMI. Among the many markers of myocardial infarction, cardiac troponin (cTn) is considered to be the most specific and longest-lasting diagnostic indicator of myocardial infarction. Among them, cTnI can show the highest peak earlier than other subtypes, and the highest peak The stability after the peak is the best, which is more important for th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68Y02A50/30
Inventor 潘宏程刘廙人
Owner GUILIN UNIVERSITY OF TECHNOLOGY
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