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Test paper for one-step detection for pig virus diarrhoea disease pathogen

A porcine viral diarrhea and test strip technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as not suitable for rapid detection or diagnosis, not easy and fast operation, multiple operation steps and experience, etc., to achieve intuitive display , Intuitive results and easy operation

Inactive Publication Date: 2009-01-07
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Virus isolation and identification and RT-PCR technology require laboratory conditions and professionals, and the operation process is cumbersome, and the detection is time-consuming and laborious; ELISA and fluorescent antibody detection methods require a microplate reader and a fluorescent microscope respectively, and require more operations. procedures and experience; moreover, these methods can only detect viral antigens for one porcine disease at a time
It can be seen that although the above methods are specific or sensitive in detection, they are not easy and fast enough to operate, and are not suitable for rapid detection or diagnosis at the grassroots level or on-site.

Method used

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  • Test paper for one-step detection for pig virus diarrhoea disease pathogen
  • Test paper for one-step detection for pig virus diarrhoea disease pathogen

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0037] (1) Preparation of test sample solution: Aseptically take fresh diarrhea feces or small intestine contents of sick pigs, make a 1:2-5 suspension with physiological saline, set it at 4°C for clarification or centrifuge at 3000r / min for 20 minutes, take the upper Clear fluid awaits inspection.

[0038] (2) Detection operation: Insert the test end of the quick detection test strip of the present invention into the clarified liquid of the sample to be tested, the insertion depth is not more than the marking line 9, take out the test strip after about 30 seconds, place it horizontally for about 1 to 5 minutes, and simultaneously Observation results.

[0039] (3) Judgment of results: If only one (one) brown-red control imprint C is displayed on the cellulose membrane of the test strip, it means that the test result is negative, indicating that TGEV, PEDV and RV were not detected in the tested sample solution If the cellulose film on the detection test strip has a brown-red c...

Embodiment 1

[0040] Example 1: One-step test strip for detecting the pathogen of porcine viral diarrhea disease

[0041] see figure 1 and figure 2 In the figure, 1 is the support layer, which is made of hard plastic sheet strips, 2 is the sample adsorption fiber layer at the test end, which is made of glass wool, and 3 is the adsorption gold-labeled antibody fiber layer, which is adsorbed with colloidal gold-labeled anti-TGEV, The glass wool of anti-PEDV and anti-RV three kinds of monoclonal antibodies, prepare its gold standard monoclonal antibody glass wool according to the preparation method described in the above-mentioned specific embodiment 3, 4 is cellulose membrane layer, adopts nitrocellulose membrane to make , 5 is a water-absorbing material layer, made of water-absorbing filter paper, each layer of numbering 2, 3, 4, and 5 is pasted on the hard plastic sheet strip 1 from the left end test end to the right, and the fibers at the junction are interpenetrated with each other. On...

Embodiment 2

[0043] Embodiment two: one step detects the test strip of porcine viral diarrhea disease pathogen, is basically the same as embodiment one, and difference is:

[0044] Adsorbed gold-labeled antibody fiber layer 3 is made of glass wool adsorbed with colloidal gold-labeled anti-TGEV, anti-PEDV and anti-RV three polyclonal antibodies, and its gold-labeled polyclonal antibody is prepared according to the preparation method described in the above-mentioned specific embodiment 3. Clone antibody glass wool; on the nitrocellulose membrane layer 4, 6 is the detection blot T, P, R "|" printed with the IgG solution of three monoclonal antibodies against TGEV, anti-PEDV and anti-RV respectively, and 7 is The control blot C "|" printed with goat (or rabbit) anti-pig IgG solution, the two blots are arranged in parallel and combined as "||||". Others including test sample preparation, operation method and result judgment are the same as the operation method in Embodiment 6 and will not be re...

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Abstract

The invention discloses a test strip which is used for testing porcine viral diarrhea pathogen in one step, and the test strip comprises a support layer, an adsorption layer and a protection layer. The adsorption layer has a sample fiber layer, a gold antibody fiber layer, a cellulose film layer and a hygroscopic material layer at the handle end sequentially from a test end. The cellulose film layer has a test blot and a contrast blot, and the gold antibody fiber layer is adhered with at least one of monoclonal antibodies or polyclonal antibodies of anti-TGEV, anti-PDEV and anti-RV with colloid gold mark; the test blot is printed by at least one of polyclonal antibody liquid or monoclonal antibody liquid of anti-TGEV, anti-PDEV and anti-RV, and the contrast blot is printed by IgG solution of sheep or rabbit anti-mouse or IgG solution of sheet or rabbit anti-porcine. When used for testing, the test strip has the advantages of strong specificity, high sensitivity, convenient and rapid operation and direct and accurate testing result. The test strip is especially suitable for fast identifying and diagnosing on site.

Description

1. Technical field: [0001] The invention relates to a device for detecting the pathogens of porcine viral infectious diseases, in particular to a test strip for detecting at least one of the three pathogens of porcine viral diarrhea diseases in one step. 2. Technical background: [0002] Porcine diarrheal disease is a kind of disease that seriously affects the development of pig industry, and viral diarrhea is especially difficult to control. Porcine epidemic diarrhea virus (PEDV, code P), porcine transmissible gastroenteritis virus (TGEV, code T) and porcine rotavirus (RV, code R) are the main viruses that cause viral diarrhea in pigs. The clinical symptoms of porcine diarrheal disease caused by pigs are similar, and there are often mixed infections in the same pig farm. Therefore, in order to prevent and treat porcine viral diarrhea in time, a specific method for rapid differential diagnosis of porcine viral diarrhea is needed, that is, rapid detection of related viruses ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577G01N33/545G01N33/544
Inventor 张改平肖治军杨艳艳李学伍邓瑞广杨继飞赵东王爱萍邢广旭柴书军刘庆堂
Owner HENAN ACAD OF AGRI SCI
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