New colloidal gold immunochromatographic method for fast qualitative and quantitative detection of ricin

A technology of ricin and colloidal gold, applied in the direction of measuring devices, analysis materials, instruments, etc., can solve the problems of expensive equipment, long cycle, cumbersome operation, etc., and achieve the effect of saving manpower and material resources

Inactive Publication Date: 2009-12-23
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of method has the characteristics of high efficiency, sensitivity, less sample usage (pmol-μpmol), automation, etc., but the disadvantages are that it requires expensive equipment, and the cycle is long and the operation is cumbersome. It is not suitable for wide application and is only suitable for experiments. room application

Method used

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  • New colloidal gold immunochromatographic method for fast qualitative and quantitative detection of ricin
  • New colloidal gold immunochromatographic method for fast qualitative and quantitative detection of ricin
  • New colloidal gold immunochromatographic method for fast qualitative and quantitative detection of ricin

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1: the preparation of colloidal gold immunochromatography test strip

[0055] Materials and methods

[0056] 1. Antigen and antibody

[0057] Rabbit anti-ricin A antibody (RTA2-G) and rabbit anti-SEB antibody were provided by the Institute of Microbial Epidemiology, Academy of Military Medical Sciences. Ricin antibody (RCA) was purchased from Sigma, USA. Goat anti-rabbit IgG (purchased from Dingguo Biotechnology Company)

[0058] 2. Consumables for chromatography test strips

[0059] Bonding pad (glass fiber), nitrocellulose membrane (NC membrane, SHF 1350225), sample pad, absorbent pad, and filter paper were purchased from Minipore.

[0060] 3. Experimental equipment

[0061] The gold standard immunoassay analyzer can be purchased from the analyzer jointly developed by Shanghai Institute of Optics and Fine Mechanics, Chinese Academy of Sciences and China Academy of Inspection and Quarantine.

[0062] 4. Preparation of colloidal gold immunochromatograph...

Embodiment 2

[0069] Embodiment 2: the judgment of qualitative and quantitative detection of sample

[0070] 1. Sample processing

[0071] Use the sample treatment solution (5mM PB pH7.4) to carry out gradient dilution of viscous liquid samples such as ham sausage, milk powder, milk, etc., add 1 / 100 (W / V) to the sample treatment solution to dissolve solid food such as ham sausage, and take the Clear, stand still and take the supernatant. Different doses of ricin were added as mock test samples.

[0072] 2 qualitative detection

[0073] Add 100ul of the treated sample and sample treatment solution (as a negative sample) to the end of the prepared chromatographic strip sample pad, let it stand for 15min, and observe the result. If both the test strip and the quality control strip appear red, it is judged as positive, and if only the quality control strip appears red, it is negative.

[0074] 3. Quantitative detection

[0075] 3.1 Determination of judgment value (CUT-OFF)

[0076] Accord...

Embodiment 3

[0079] Embodiment 3: sensitivity test

[0080] 1. Sensitivity of quantitative detection

[0081] Detecting different concentrations of ricin, the calculated judgment value (CUT-OFF) is 0.037, and the average T / C ratio of the ricin at a concentration of 0.925 μg / ml is positive if the value is higher than 0.037; When the toxin concentration is greater than 103cfu / ml, the average T / C ratio is greater than 0.037, so the quantitative detection sensitivity is 0.925μg / ml.

[0082]Table 1 Detection results of various concentrations of ricin colloidal gold immunochromatographic test strips

[0083]

[0084] 2. Intuitive detection sensitivity evaluation

[0085] Colloidal gold immunochromatographic test strips were prepared according to the determined optimal reaction conditions to detect different concentrations of ricin. The ricin was diluted with the sample treatment solution to a concentration of 14.8 μg / ml, 7.4 μg / ml, 3.7 μg / ml, 1.85 μg / ml, 0.925 μg / ml, and 0.4625 μg / ml, and ...

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Abstract

The invention establishes a fast detection method of ricin by utilizing immuno-chromatographic technology of colloidal gold labeling and double antibody sandwich. The established colloidal gold immunochromatographic method for detecting ricin can detect the ricin in a sample fast, sensitively, specifically and accurately, can realize quantitation and is applicable to field fast detection.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a rapid qualitative and quantitative detection method for ricin and a colloidal gold immunological detection test strip. Background technique [0002] Ricin toxin (RT) is a plant protein toxin extracted from the endosperm of castor seeds. Because RT has a wide range of sources and strong toxicity, it has been studied as a weaponized toxin by foreign troops. The Convention on the Prohibition of Chemical and Biological Weapons lists RT as one of the most stringent controls. Since the "9.11" terrorist attack in the United States, as many terrorist organizations and extremists developed, possessed and carried RT, they appeared in London, the capital of the United Kingdom, making this deadly toxin the most likely to be used as a biological toxin for terrorist attacks. aroused widespread international attention. In response to bioterrorism, it is of great significance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/552G01N33/532G01N33/558
Inventor 王静王景林聂聪杨宇孙肖红高姗胡孔新张晓龙
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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