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Immunity chromatography test paper for detecting ricin and preparation method thereof

An immunochromatographic test paper, ricin technology, applied in the directions of measuring devices, analytical materials, instruments, etc., can solve problems such as differences in the sensitivity of ricin, and the detection of ricin by immunochromatographic test paper has not yet been found, and achieve sample processing. simple effect

Inactive Publication Date: 2008-06-25
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the sensitivity of different detection methods to detect ricin varies
There is no report on the detection of ricin by immunochromatographic test paper prepared by immunocolloidal gold technology

Method used

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  • Immunity chromatography test paper for detecting ricin and preparation method thereof
  • Immunity chromatography test paper for detecting ricin and preparation method thereof
  • Immunity chromatography test paper for detecting ricin and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of immunochromatographic test paper for detecting ricin

[0026] 1. Material

[0027] Chloroauric acid (HAuCl 4 ·4H 2 O), analytically pure, Shanghai No. 1 Reagent Factory;

[0028] Trisodium Citrate (Na 3 C 6 H 5 O 7 ·2H 2 O), analytical pure, Beijing Chemical Plant;

[0029] Ricin, prepared by our laboratory according to conventional purification methods;

[0030] Bovine serum albumin (BSA), analytically pure, imported aliquot;

[0031] Polyethylene glycol (PEG), analytically pure, imported subpackage;

[0032] Nitrocellulose membrane, purchased from Millipore Corporation, HFB 504;

[0033] Glass fiber membrane, purchased from Millipore, USA;

[0034] Recombinant Ricin B-chain Protein [9] : Using codon optimization software to re-optimize the design of RTB coding gene sequence, use 18 long-fragment oligonucleotide primers to synthesize RTB gene fragment by two-step overlapping PCR, and introduce 6×His tag sequence into the C-terminal of RTB, insert a...

Embodiment 2

[0054] Example 2 Ricin detection test

[0055] 1. The sensitivity of ICA test strips to detect ricin

[0056] Ricin was purified and prepared by the State Key Laboratory of Pathogenic Microbial Biosafety, Institute of Microbial Epidemiology, Academy of Military Medical Sciences [11] As a standard, after limited dilution with physiological saline, as the sample test solution, take the ricin ICA test paper and immerse it in the test solution of different ricin concentration samples. The results will be observed after 5 minutes, and the observation will stop after 15 minutes. Results report: 1 red (control) precipitation line is negative, that is, no ricin is detected; 2 red (sample and control) precipitation lines are positive, that is, ricin is detected.

[0057] The ricin ICA test strip provided by the present invention can detect 20ng / ml. It is reported in the literature that the sensitivity of the ELISA method to detect pure ricin is 100pg / mL.

[0058] 2. ICA test paper for dete...

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Abstract

The invention discloses an immunity chromatographic test paper for detecting ricin and a preparation method thereof. The immunity chromatographic test paper for detecting the ricin provided by the invention comprises an absorbent paper washer, a fiber membrane which is closely connected to the absorbent paper washer and coated with anti-ricin A-chain protein antibodies, a gold conjugate pad which is closely connected on the fiber membrane, consists of anti-ricin B-chain protein antibodies and is immunized from gold colloid probes, and a sample washer which is closely connected to the gold conjugate pad. Moreover, a backboard is also closely connected under the absorbent paper washer for convenience in use. By adoption of the immunity chromatographic test paper for detecting the ricin, laypeople can operate according to specifications; treatment processes of specimen is simple, and results can be observed within 5 minutes; at least 20 nanogram per milliliter ricin can be detected, and detection sensibility is not affected by specimen sources. The ricin immunity detection test paper has the advantages of simplicity, convenience, sensitivity, specificity and quickness and is suitable for clinical and field use.

Description

Technical field: [0001] The invention relates to a ricin test paper and a preparation method thereof. Background technique: [0002] Ricin toxin (RT) is a plant protein toxin extracted from the endosperm of castor bean seeds. Its molecular weight is about 64kDa. The two polypeptide chains are called RTA chain (Ricin toxin A, RTA) and RTB chain ( Ricin toxin B, RTB). RTA is an active chain with a molecular weight of approximately 32 kDa. RTB is a combined chain with a molecular weight of about 34 kDa, and the two are connected by disulfide bonds. Because ricin has a wide range of sources and high toxicity, it has been studied by foreign forces as a weaponized toxin. The Convention on the Prohibition of Chemical and Biological Weapons lists ricin as one of the most stringent targets. Especially since the "9.11" terrorist attacks in the United States, as many terrorist organizations and extremists developed, possessed and carried ricin toxin, the British capital London was discovere...

Claims

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Application Information

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IPC IPC(8): G01N33/549G01N33/577G01N33/532
Inventor 王景林王俊虹康琳高姗
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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