Fluorescence immunoassay kit used for detecting porcine circovirus type 2

A porcine circovirus and fluorescent immunity technology, applied in fluorescence/phosphorescence, measuring devices, instruments, etc., can solve the problems of high technical requirements, long time-consuming diagnosis, high cost, and achieve good repeatability, fast detection speed, specificity strong effect

Active Publication Date: 2013-12-25
YEBIO BIOENG OF QINGDAO
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  • Abstract
  • Description
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Problems solved by technology

Among them, PCR and ISH require a clear and common nucleic acid sequence for identification, which requires high technical requirements for the experimenter and is prone to false positives; IHC has high requirements for the operator’s pathological section technology, and the detection procedure is relatively cumbersome; and the animal regression experiment requires feeding experiments. Observing the clinical symptoms of animals waiting for the disease is used for identification, so the diagnosis takes a long time and the cost is high; the ELISA antibody detection kit can only detect PCV2 antibodies, and cannot determine whether the pigs are infected with PCV2

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  • Fluorescence immunoassay kit used for detecting porcine circovirus type 2
  • Fluorescence immunoassay kit used for detecting porcine circovirus type 2
  • Fluorescence immunoassay kit used for detecting porcine circovirus type 2

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Embodiment Construction

[0015] The present invention will be described in detail below in conjunction with specific examples. The chemicals or reagents used in the present invention can be selected from existing other types of products according to their functions, and are not limited to the specific records of the examples. For experimental methods that do not indicate specific conditions in the examples, conventional conditions can usually be used, such as the conditions described in the "Molecular Cloning Experiment Guide" written by J. Sambrook (Sambrook), or as recommended by the manufacturer. conditional run.

[0016] (1) Preparation of single-factor serum antibody (primary antibody) for detection of PCV2

[0017] 1. Preparation of the Cap protein of PCV2 whose amino acid sequence is SEQ ID NO:2

[0018] According to a sequence at both ends of the PCV2ORF2 gene and the multiple cloning restriction site of the expression vector PET-28a, the specific primer PCV2ORF2-F of PCV2ORF2 was designed: 5...

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Abstract

The invention provides a fluorescence immunoassay kit used for detecting porcine circovirus type 2 (PCV2), which comprises a single factor serum antibody used for detecting PCV2, an FITC (fluorescein isothiocyanate) labeled goat anti-mouse IgG (immunoglobulin G), a positive control sample, stationary liquid and PBS (phosphate buffer) washing liquor, wherein the single factor serum antibody used for detecting PCV2 is prepared from a PCV2 Cap protein antigen immunized mouse with an amino acid sequence of SEQ ID NO: 1. The main ingredient in the kit used for detecting PCV2 prepared in the invention is a single factor serum antibody of mouse anti-porcine circovirus type 2 and is prepared from the antigen immunized mouse with the amino acid sequence of SEQ ID NO: 1, and the prepared antibody can generate a specific reaction with PCV2. The fluorescence immunoassay kit of porcine circovirus type 2 created on the basis of the single factor serum can be used for detecting antigen in the diseased tissues of an PCV2 infected pig on clinic and judging whether the pig is infected with PCV2, and the fluorescence immunoassay kit is strong in specificity, high in sensitivity, good in repeatability and high in detection speed.

Description

technical field [0001] The invention belongs to the technical field of livestock virus detection, and in particular relates to a fluorescent immunoassay kit for detecting porcine circovirus type 2. Background technique [0002] In 1991, John Harding of Canada reported Postweaning Multisystemic Wasting Syndrome (PMWS), and after extensive and in-depth investigation and research, it was confirmed in 1998 that porcine circovirus (Porcine Circovirus, PCV) was the main pathogen. Existing studies have shown that porcine circovirus disease (Porcine Circovirus Disease, PCVD) is a clinical symptom caused by PCV2 infection, which is progressive weight loss, shortness of breath, anemia, diarrhea, jaundice, interstitial pneumonia, lymphadenitis And nephritis, mainly against weaned piglets, and PMWS, porcine dermatitis and nephritis syndrome (Porcine Dermatitis and Nephropathy Syndrome, PDNS), porcine respiratory syndrome (Porcine Respiratory Disease Complex, PRDC), A2 congenital tremor ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N21/64
Inventor 张恒邹敏范根成刘蕾徐保娟胡潇申洪银陶晓珊
Owner YEBIO BIOENG OF QINGDAO
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