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Double-antibody sandwich ELISA kit for detecting porcine acute diarrhea syndrome coronavirus

A technology for acute diarrhea and coronavirus, applied in the field of double-antibody sandwich ELISA kits, can solve the problems of unsuitability for rapid detection, expensive equipment, long test cycle, etc., and achieve rapid detection methods and monitoring methods with good specificity

Active Publication Date: 2022-02-25
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology describes an improved way to detect or monitor viruses like coronavirus (which causes COVID-19). It involves capturing these particles into tiny clumps called pseudo cages that are then immobilized onto solid surfaces such as glass plates. These false cage structures have unique properties which make them more effective at identifying small amounts of harmful agents compared to other techniques like enzyme linked immunosorbant assay(ELISA) tests. By comparing different types of molecules captured on this surface against each others's ones, it becomes possible to identify new strains associated with diseases caused by certain bacteria.

Problems solved by technology

This patents discuss how different types of viral agents can cause sickness from sialidacrovirus that causes AIDS called Acquired Immune Deficiency Syndrom ("SIRS"), while another type called Porcoletype VCuilleminor ("PTCV") affects only humans who get affected with this illness later during childhood. These two types may lead to secondary bacteria like Salmonella enteritica serovars causing deaths if left untreatable through traditional ways like drinking water containing harmful germ Clostrids.

Method used

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  • Double-antibody sandwich ELISA kit for detecting porcine acute diarrhea syndrome coronavirus
  • Double-antibody sandwich ELISA kit for detecting porcine acute diarrhea syndrome coronavirus
  • Double-antibody sandwich ELISA kit for detecting porcine acute diarrhea syndrome coronavirus

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Embodiment Construction

[0056] In the following, the present invention will be described in more detail through specific embodiments, so as to facilitate the understanding of the technical solution of the present invention, but it is not used to limit the protection scope of the present invention.

[0057] The SADS-CoV GDS04 strain was donated by Professor Cao Yongchang of Sun Yat-sen University.

[0058] 1. Construction, expression and purification of SADS-CoVN protein recombinant plasmid

[0059] 1.1 Construction of pET32a-N recombinant plasmid

[0060] Using the N protein of SADS-CoV GDS04 strain (Genbank accession number: MF167434.1) as a reference, design specific primers, upstream P1: 5′-CGC GGATCC ATGGCCACTGTTAATTGG-3′, downstream P2: 5′-CCG CTCGAG CTAATTAATAATCTCATC-3', BamHI and XhoI restriction sites (underlined part) were introduced at the 5' ends of the upstream and downstream primers. After the primers were synthesized, PCR amplification was performed using the SADS-CoV GDS04 strain...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a double-antibody sandwich ELISA kit for detecting porcine acute diarrhea syndrome coronavirus. The double-antibody sandwich ELISA kit for detecting the porcine acute diarrhea syndrome coronavirus comprises an ELISA plate coated with a rabbit anti-SADS-CoV N protein polyclonal antibody, and an HRP (horse radish peroxidase) labeled mouse anti-SADS-CoV N protein monoclonal antibody. The kit further comprises an enzyme-labeled secondary antibody, a confining liquid, a diluent, a washing liquid, a developing liquid and a stop solution. The double-antibody sandwich ELISA kit established by the invention has relatively good specificity, sensitivity and intra-batch and inter-batch repeatability. The lowest virus detection amount of the kit is 10-4.42TCID50/0.1 mL, the kit does not react with PEDV, TGEV and PDCoV, and the intra-batch and inter-batch repeated variable coefficient is less than 10%. Compared with an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) method, the detection coincidence rate of the double-antibody sandwich ELISA kit disclosed by the invention is 93.93%. The SADS-CoV antigen double-antibody sandwich ELISA detection method established by the invention has good sensitivity, specificity and repeatability, and can be used for clinical detection of SADS-CoV.

Description

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Claims

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Application Information

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Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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