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Immunofluorescence detection test strip for rapidly detecting oyster herpesvirus antigen and application of immunofluorescence detection test strip

An oyster herpes virus and immunofluorescence technology, applied in the field of immunofluorescence detection test strips, can solve the problems of high professional level requirements of detection operators, inability to achieve rapid detection, and contamination of positive samples, and achieve fast detection speed and detection cost. Low, simple to prepare effect

Pending Publication Date: 2022-03-04
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection of OsHV-1 is based on the detection of the level of viral nucleic acid, which requires a high level of professionalism for the detection operators; the detection field requires supporting equipment, reagent storage space, and needs to maintain a high degree of cleanliness to avoid positive sample contamination
At present, the processing cycle based on nucleic acid detection technology is long, and the purpose of rapid on-site detection cannot be achieved

Method used

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  • Immunofluorescence detection test strip for rapidly detecting oyster herpesvirus antigen and application of immunofluorescence detection test strip
  • Immunofluorescence detection test strip for rapidly detecting oyster herpesvirus antigen and application of immunofluorescence detection test strip
  • Immunofluorescence detection test strip for rapidly detecting oyster herpesvirus antigen and application of immunofluorescence detection test strip

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Experimental program
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Embodiment 1

[0035] Embodiment 1: Preparation of OsHV-1 immunofluorescence detection test strip

[0036] 1. Prokaryotic expression and purification of EPI protein

[0037] 1) According to the France 2013 strain sequence published by GenBank (GenBank: NC_005881.2), using the pUC57 plasmid as a framework, a recombinant vector containing the OsHV-1 EPI gene (SEQ ID NO.3) was synthesized by Shanghai Sangon Bioengineering Co., Ltd. , and named as pUC57-EPI respectively;

[0038] 2) Using the above-mentioned synthetic recombinant vector pUC57-EPI as a template, amplify and clone EPI fragments into prokaryotic expression vectors pGEX-4t-1 and pET28a-SUMO, respectively, and construct prokaryotic expression vectors pGEX-4t-1-EPI and pET28a-EPI;

[0039] 4) The prokaryotic expression vectors pGEX-4t-1-EPI and pET28a-p25 were transformed into Escherichia coli BL21(DE3) competent cells, respectively, and the expression strains BL21-pGEX-4t-1-EPI and BL21-pET28a-EPI were obtained;

[0040] 5) Express...

Embodiment 2

[0062] Example 2: Rapid detection of OsHV-1 antigen using test strips

[0063] 1. Rapid qualitative detection of OsHV-1 antigen

[0064] 1.1 Preparation of detection sample solution

[0065] The shellfish mantle tissue to be tested is collected, and an appropriate amount of tissue lysate is added for grinding and crushing.

[0066] 1.2 Sample testing

[0067] Drop the sample solution to be tested onto the sample pad of the test strip, and place it horizontally for 3-5 minutes to observe the results.

[0068] 1.3 Judgment of test results

[0069] Two fluorescent bands appear on the test strip test pad (that is, both the test line and the quality control line are colored), indicating that the sample to be tested contains OsHV-1 antigen; only one fluorescent band (quality control line) is negative, Indicates that no OsHV-1 antigen was detected in the sample to be tested; no bands appeared on the test strip, indicating improper detection operation or failure of the test strip....

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Abstract

The invention belongs to the technical field of biological detection, and particularly relates to an immunofluorescence detection test strip for rapidly, qualitatively and quantitatively detecting an OsHV-1 (Ostreid Herpes Virus 1) antigen and application of the immunofluorescence detection test strip. The immunofluorescence test strip provided by the invention takes the ORF104 protein as shown in SEQ ID NO.1 as an antigen molecule to be detected. The operation steps are simple, the detection speed is high, the result can be qualitatively observed and quantitatively measured, and the detection cost is low. The to-be-detected sample is simple to prepare and only needs to be matched with a tissue lysis solution to crush the tissue, and a tedious nucleic acid extraction process is not needed.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to an immunofluorescence detection test strip for rapid qualitative and quantitative detection of Ostreidherpesvirus (Ostreidherpesvirus1, OsHV-1) antigen and an application thereof. Background technique [0002] Oyster herpes virus (OsHV-1) belongs to the order Herpesvirales and the family Malacoherpesviridae, and its host range involves many cultured bivalve molluscs in my country, including oysters, scallops and cockles. Oyster herpes virus can cause mass mortality in shellfish populations after infecting farmed shellfish populations. Since the bivalve mollusc culture mode belongs to open water body culture, it is difficult to apply pesticides to treat the disease of cultured molluscs, so the risk control of pathogens during the culture process is more important. In order to realize the regular monitoring task of oyster herpes virus, a pathogen, it is ur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/558G01N33/543G01N33/533
CPCG01N33/56994G01N33/558G01N33/54313G01N33/533G01N2333/03
Inventor 辛鲁生于江南白昌明王崇明
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI