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A kind of phospholipase a2 lateral chromatography detection test strip, detection card and application thereof
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A technology of lateral chromatography and test strip detection, which is applied in the biological field and can solve problems such as increased operational errors, high cost of test strip production, and low sensitivity
Active Publication Date: 2022-03-18
GUANGDONG NO 2 PROVINCIAL PEOPLES HOSPITAL
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Although the two-step analysis solves the problem of low sensitivity caused by the weak binding ability between the signal probe and the analyte, the two-step operation increases the complexity of the operation, increases the possibility of operational errors, and makes the entire detection system more cumbersome, Test strips are more expensive to make
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[0058] 1. Preparation of phospholipase A2 lateral flow detection card
[0059] (1) Preparation of sample pad:
[0060] Prepare the sample pad treatment solution. The specific formula is shown in Table 1-1 below. Soak the glass fiber membrane in the sample pad treatment solution for 10 minutes, and then place the soaked glass fiber membrane in a 37°C drying oven to dry overnight. The glass fiber membrane is cut into 12mm strips, which is the sample pad.
[0061] Table 1-1
[0062]
[0063] The above-mentioned components were respectively added into 100 mL of 0.015 mol / L PBS (BOSTER, PBS-AR0030) buffer solution with pH=7.4 and dissolved to obtain the final product.
[0064] (2) Preparation of binding pad:
[0065] 1) Signal probe preparation: In this scheme, time-resolved fluorescent microspheres (EuNPs, particle size 190nm, solid content 1mg / mL, excitation wavelength 365nm, emission wavelen...
Embodiment 2
[0090] The detection of embodiment 2 phospholipase A2
[0091] (1) Standard testing
[0092] At the same time, the standard solution of different concentrations (0, 10, 50, 100, 250, 500, 1000 ng / mL, Yuanye Biology, S25308) was detected with the three detection cards prepared above, and each was repeated 3 times. The detection methods are as follows:
[0093] Liquid flow control lateral immunochromatography test card operation: Add the sample to be tested into the sample hole of the test card, unplug the flow barrier after 5 minutes, and use the fluorescence reader to record the fluorescence signals of the test line and quality control line after 5 minutes.
[0094] Conventional lateral flow immunochromatography test card operation: Add the sample to be tested into the sample hole of the test card, and use a fluorescence reader to record the fluorescence signals of the test line and quality control line after 10 minutes.
[0095] The operation of the immunochromatography tes...
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Abstract
The invention relates to a phospholipase A2 lateral chromatography detection test strip, a detection card and its application. By adding a baffle between the binding pad and the chromatographic membrane in the test strip, the flow rate of the sample on the binding pad can be controlled. flow time; and on this basis, a phospholipase A2 lateral chromatography detection card is designed, the housing is provided with a flow control switch, and the movable end of the baffle is controlled by the flow control switch on the housing. The liquid flow control switch can start the subsequent analysis process after the recognition reaction between the fluorescent microsphere-labeled antibody and the analyte is completed. It is found through experiments that the phospholipase A2 detection card designed by the lateral chromatography device of the present invention has a lower detection limit and a stronger fluorescent signal than the conventional lateral immunochromatographic detection card, which indicates that it has broad clinical application prospects .
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a phospholipase A2 lateral chromatography detection test strip, a detection card and applications thereof. Background technique [0002] Phospholipase A2 (phospholipase A2, PLA2) is a hydrolase that can catalyze the two-position acyl group on the phospholipid glycerol molecule. Rate-limiting enzymes, produced by lipid mediators, play a key role in membrane channel activation, information transmission, hemodynamics, and pathophysiological processes during inflammation and tissue injury, as well as in the regulation of intracellular and extracellular metabolism. PLA2 activity is elevated in serum of patients with acute pancreatitis, septic shock, trauma and multiple organ failure (MSOF). In multiple trauma, changes in PLA2 activity may be an early signaling parameter in the process of tissue damage in MSOF. [0003] Lateral flow assay strip (LFAS) is a microporous chromatographic mem...
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