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New colloidal gold immunochromatographic method for fast quantifying and detecting Listeria monocytogenes and colloidal gold immunological detection test strip

A technology of Listeria monocytogenes and colloidal gold, which is applied in the rapid qualitative and quantitative detection of Listeria monocytogenes and in the field of colloidal gold immunoassay test strips, can solve the problem of fewer products detected in the field of food hygiene, and achieve Good stability and accurate detection effect

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

In recent years, this method has developed rapidly and has been widely used in the field of biomedicine, especially in medical testing, but there are few products used in the field of food hygiene testing.

Method used

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  • New colloidal gold immunochromatographic method for fast quantifying and detecting Listeria monocytogenes and colloidal gold immunological detection test strip
  • New colloidal gold immunochromatographic method for fast quantifying and detecting Listeria monocytogenes and colloidal gold immunological detection test strip
  • New colloidal gold immunochromatographic method for fast quantifying and detecting Listeria monocytogenes and colloidal gold immunological detection test strip

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: the preparation of colloidal gold immunochromatography test strip

[0032] 1. Antigen and antibody

[0033] Listeria monocytogenes is Listeria monocytogenes, bacteria number 19111, from the Institute for the Control of Pharmaceutical and Biological Products of the Ministry of Health,

[0034] Listeria monocytogenes P60 polyantibody is Listeria monocytogenes, multiplecloning antibody of P60 comes from Jilin Inspection and Quarantine Bureau),

[0035] Goat anti-rabbit IgG can be purchased from Dingguo Biotechnology Company.

[0036] 2. Consumables for chromatography test strips

[0037]Bonding pad (glass fiber), nitrocellulose membrane (NC membrane, SHF 1350225), sample pad, absorbent pad, and filter paper were purchased from Minipore.

[0038] 3. Experimental equipment

[0039] Gold Standard Immunity Analyzer (jointly developed by Shanghai Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, and China Academy of Inspection and Quarantine...

Embodiment 2

[0047] Embodiment 2: Qualitative and quantitative detection of samples

[0048] 1. Sample processing

[0049] 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 Listeria monocytogenes were added as mock test samples.

[0050] 2. Qualitative detection

[0051] 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.

[0052] 3. Quantitative detection

[0053] 3.1. Determination of judgment value

[0054] According to...

Embodiment 3

[0057] Embodiment 3: sensitivity test

[0058] 1. Sensitivity of quantitative detection

[0059] Detect different concentrations of Listeria monocytogenes, the calculated judgment value (CUT-OFF) is 0.017, the concentration is 10 1 cfu / ml, 10 2 The average T / C ratio of cfu / ml Listeria monocytogenes is 0.00, which is negative if it is lower than 0.017; 10 3 cfu / ml~10 8 The average T / C ratios of cfu / ml Listeria monocytogenes were all higher than 0.017 and the results were positive; when the concentration of Listeria monocytogenes was greater than 10 3 When cfu / ml, the average value of T / C ratio is greater than 0.017, so the quantitative detection sensitivity is 10 3 cfu / ml.

[0060] Table 1 Detection results of various concentrations of Listeria monocytogenes colloidal gold immunochromatographic test strips

[0061]

[0062]

[0063] 2. Intuitive detection sensitivity

[0064] Colloidal gold immunochromatographic test strips were prepared according to the determined...

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Abstract

The invention establishes a colloidal gold immunochromatographic method for detecting Listeria monocytogenes, the method can detect Listeria monocytogenes in a sample fast, sensitively, specifically and accurately, and can realize quantitation and is applicable to field fast detection. The invention belongs to the field of biological detection, and particularly relates to a method for fast qualitative detection and quantitative detection of Listeria monocytogenes and a colloidal gold immunological detection test strip.

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 the detection of Listeria monocytogenes and a colloidal gold immunological detection test strip. Background technique [0002] Listeria monocytogenes (Listeria monocytogenes, Listeria monocytogenes), is the most important human food-borne pathogenic bacteria in Listeria (Listeria), is also a kind of zoonotic pathogenic bacteria, belongs to Listeria It can cause meningitis, encephalitis, sepsis, endocarditis, miscarriage, stillbirth and abscess in humans and animals, and the mortality rate of the patients can reach 30%-40%. In recent years, many countries have reported food poisoning incidents caused by contamination of Listeria monocytogenes. Therefore, government departments around the world are paying more and more attention to food poisoning caused by Listeria monocytogenes, and have formulated some new ...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/577G01N33/558G01N33/551
Inventor 王静王振国谢士嘉杨宇姚李四孙肖红胡孔新张晓龙
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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