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A ricin and detection line technology, which is applied in the field of rapid quantitative detection card for ricin, can solve the problems of large volume, low sensitivity and complicated operation of detection equipment, and achieve improved sensitivity and precision, high detection limit and low cost Effect
Pending Publication Date: 2021-06-11
ACADEMY OF MILITARY MEDICAL SCI +1
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At present, the detection methods for RT mainly include: ELISA method is the most classic method in the world, and its sensitivity can reach ng / mL, but its application is relatively limited due to complicated operation, large detection equipment, and long time consumption; colloidal gold immunochromatography test paper Strips use the visual positioning properties of colloidal gold labeling technology to get rid of the limitations of special instruments and equipment, and are low in cost and easy to operate. The disadvantage is that it can only achieve semi-quantitative detection and low sensitivity.
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preparation example Construction
[0056] (1) Preparation of ricin:
[0057] 100g castor seeds, soaked, homogenous, centrifugal, etc., then precipitate all proteins, followed by dialysis, centrifugal, taking 2 mL of the supernatant, adding medium galactoside agarose gel, slowly adding the column pipe, Then eluted protein (containing RT and its lectin) with D-galactose disposable. The above-mentioned elura was concentrated with PEG 20000 and purified with Sephacryl S-100 to purify the gel column to obtain ricin. Castor toxin affinity purified SDS-PAGE electrophoresis figure 1 . Ricin SDS-PAGE Electrophoresis figure 2
[0058] Identification of ricin
[0059] Purified RT line immunoblot analysis (Western blot, WB), then cut an electrophoretic gel containing the target strip, transfer the protein strip to the nitrocellulose film (NC film) by semi-dry membrane instrument On, 1mA / cm 2 The power transfer is transferred for 2 h; then, the NC film is placed in the closed liquid in the closed night, and the washing liquid...
Embodiment 1
[0064] Example 1, a castor spin detection card is prepared
[0065] Castorin detection principle (double antibody sandwich method): figure 1 The detection card shown, 6 packs of the NC membrane control line 6 packs of the sheep anti-chicken IGE, the detection line 7 packs are placed in the lanthanum pad in the lanthanum pad, lanthanum, lanthanum, lanthanum Aim.
[0066]Step 1. Paste the NC film (Sartorius CN140) on the PVC bottom plate, and sprayed to 0.3 mg / ml of sheep anti-chicken IGG to form a quality control line (C wire) on the NC film. And the castor toxin polyclonal antibody has been diluted to 2 mg / ml, the spray mass was 1 μl / cm, and then treated at a temperature of 37 ° C for 16 hours;
[0067] Step Second, preparing lanthanoid fluorescent microspheres with chicken IGY and lanthanum fluorescent microspheres labeled with ricin monoxin. 1 ml of lanthanum fluorescence microspheres were added to 5 mL of MES (2- (N-morphine) acenesulfonic acid) buffer (0.05 m, pH 7.2), a...
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Abstract
The invention discloses a double-hole rapid quantitative detection card for ricin. The double-hole rapid quantitative detection card comprises a PVC bottom plate, a water absorption pad, a nitrocellulose membrane, a marker pad, a sample pad and a diluent pad which are arranged in a shell; the nitrocellulose membrane is fixed on the PVC bottom plate; the left end of the nitrocellulose membrane is in lap joint with the water absorption pad; the right end of the nitrocellulose membrane is in lap joint with the dilution pad; the marker pad is arranged on the right side of the middle of the nitrocellulose membrane; the sample pad is arranged on the marker pad; and a quality control line and a detection line are arranged between the water absorption pad and the marker pad on the nitrocellulose membrane; window holes corresponding to the detection line and the quality control line are formed in the upper case of the shell; quantitative identification lines are arranged on the detection line and the edge of the window hole close to the marker pad; a sample adding hole corresponding to the sample pad is formed; and a cleaning hole corresponding to the dilution pad is formed. According to the detection card of the invention, a chromatographic immunoassay method of adding a sample into the sample adding hole, adding a diluent into the cleaning hole and controlling the liquid flow amount of the sample side through the quantitative marking line is adopted, so that the sensitivity and the precision of a detected object are improved, and the characteristics of rapidness, simplicity and convenience of the rapid diagnosis detection method are retained.
Description
Technical field [0001] The present invention belongs to the field of biological detection, and specifically, a rapid quantitative detection card for ricinotoxins. Background technique [0002] Ricin Toxin, RT) is a high toxic glycoprotein in castor seeds, and the content is about 1.0 to 5.0% of the grain. As an important oil crop, castor is widely distributed and planted in my country, my country's ramie seed production ranks second in the world. And RT is widely included in the "Prohibition of Chemical and Biological Weapons Convention", and is listed as Class B Biological Act (CDC) by the US CDC (CDC) . In addition, according to the data of my country's National CDC, every year, there are castor poisoning and death incidents caused by crystallic castor seeds. Therefore, in order to cope with the potential threat of castor toxins, relevant organizations of countries have been working on the prevention and control measures of castor toxins. [0003] Tlemperm treatment can only be...
Claims
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Application Information
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