PE20191840A1 - DIFFERENTIAL AND RAPID DIAGNOSTIC KIT FOR DETECTION OF INFECTIOUS CORIZA AND SWOLL HEAD SYNDROME IN THE POULTRY INDUSTRY - Google Patents

DIFFERENTIAL AND RAPID DIAGNOSTIC KIT FOR DETECTION OF INFECTIOUS CORIZA AND SWOLL HEAD SYNDROME IN THE POULTRY INDUSTRY

Info

Publication number
PE20191840A1
PE20191840A1 PE2018001160A PE2018001160A PE20191840A1 PE 20191840 A1 PE20191840 A1 PE 20191840A1 PE 2018001160 A PE2018001160 A PE 2018001160A PE 2018001160 A PE2018001160 A PE 2018001160A PE 20191840 A1 PE20191840 A1 PE 20191840A1
Authority
PE
Peru
Prior art keywords
avibacterium paragallinarum
peptides
ampv
prototype
detection
Prior art date
Application number
PE2018001160A
Other languages
Spanish (es)
Inventor
Diaz Manolo Clemente Fernandez
Eguis Jorge Eduardo Bendezu
Ruiz Sandra Steffany Morales
Original Assignee
Farm Veterinarios S A C
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Farm Veterinarios S A C filed Critical Farm Veterinarios S A C
Priority to PE2018001160A priority Critical patent/PE20191840A1/en
Priority to PCT/PE2019/000006 priority patent/WO2019245392A1/en
Priority to ARP190101668A priority patent/AR119649A1/en
Publication of PE20191840A1 publication Critical patent/PE20191840A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/12—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Hematology (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Urology & Nephrology (AREA)
  • Food Science & Technology (AREA)
  • General Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biotechnology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • Pathology (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

Con la finalidad de reducir los tiempos en el diagnostico de la coriza infecciosa (CI) y el sindrome de cabeza hinchada (SCH), enfermedades que provocan grandes perdidas en la industria avicola, el presente trabajo busco desarrollar Tiras inmunocromatograficas (TIC) que permiten detectar los agentes que provocan dichas enfermedades (Avibacterium paragallinarum y Metapneumovirus aviar(aMPV)). Para esto el trabajo se dividio en 4 etapas: 1) Bioinformatica: Se identificaron proteinas candidatas en base a la informacion genomica disponible y se identificaron peptidos inmunogenos (14 aminoacidos de longitud); 2) Produccion de Policlonales: Se sintetizaron los peptidos seleccionados y se emplearon para inmunizar conejos (New-Zeland) para luego obtener anticuerpos policlonales (AcP) y estos fueron evaluados en formatos de western blot (WB) y ELISA frente a derivados (lisados y sobrenadantes) de Avibacterium paragallinarum. 3) Produccion de Monoclonales: se continuo con aquellos peptidos inmunogenos que dieron origen AcP con una correcta reactividad y especificidad, dichos peptidos fueron empleados para inmunizar ratones BALB/C, luego se les extrajo el bazo para los ensayos de obtencion de hibridomas, seleccion de clonas productoras y purificacion de anticuerpos monoclonales. Estos anticuerpos monoclonales (Am) fueron evaluados en formatos de WB y ELISA. 4) Desarrollo de un prototipo del kit: Se diseno un set de TICs (Kit) con aquellos Am que cumplieron los criterios de evaluacion en la etapa 3, se evaluo este prototipo frente a muestras de Avibacterium paragallinarum, aMPV y otros patogenos aviares en condiciones de laboratorio. Se obtuvieron los siguientes resultados (R) para cada etapa: R1) En Avibacterium paragallinarum se identifico la proteina tonB-dT y se ubicaron 6 peptidos. R2) Avibacterium paragallinarum: De los 6, 2 peptidos produjeron anticuerpos policlonales (AcP) que identificaron correctamente la proteina tonB-dT en muestras de Avibacterium paragallinarum; R3) En Avibacterium paragallinarum: Con los 2 peptidos se obtuvieron 4 clonas de hibridomas productoras de anticuerpos monoclonales (Am); sin embargo, solo una clona (1G7G8) fue reactiva a tonB-dT. R4) Con el Am-1G7G8 se desarrollo un prototipo TIC cualitativo (presencia y ausencia) que demostro ser especifico para Avibacterium paragallinarum frente a muestras de aMPV y otros patogenos aviares. Se logro desarrollar un prototipo de TIC para la deteccion de Avibacterium paragallinarum sin presentar reactividad cruzada frente a aMPV y brinda resultados en 30 minutos de aplicada la muestraIn order to reduce the time in the diagnosis of infectious coryza (IC) and swollen head syndrome (SCH), diseases that cause great losses in the poultry industry, the present work seeks to develop immunochromatographic strips (TIC) that allow the detection of the agents that cause these diseases (Avibacterium paragallinarum and Avian Metapneumovirus (aMPV)). For this, the work was divided into 4 stages: 1) Bioinformatics: Candidate proteins were identified based on available genomic information and immunogenic peptides (14 amino acids long) were identified; 2) Production of Polyclonal: The selected peptides were synthesized and used to immunize rabbits (New-Zeland) to then obtain polyclonal antibodies (AcP) and these were evaluated in western blot (WB) and ELISA formats against derivatives (lysates and supernatants) of Avibacterium paragallinarum. 3) Production of Monoclonals: it was continued with those immunogenic peptides that gave AcP origin with a correct reactivity and specificity, said peptides were used to immunize BALB / C mice, then the spleen was extracted for the hybridoma obtaining tests, selection of Producing clones and purification of monoclonal antibodies. These monoclonal antibodies (Am) were evaluated in WB and ELISA formats. 4) Development of a kit prototype: A set of TICs (Kit) was designed with those Am that met the evaluation criteria in stage 3, this prototype was evaluated against samples of Avibacterium paragallinarum, aMPV and other avian pathogens under conditions from laboratory. The following results (R) were obtained for each stage: R1) In Avibacterium paragallinarum the tonB-dT protein was identified and 6 peptides were located. R2) Avibacterium paragallinarum: Of the 6, 2 peptides produced polyclonal antibodies (AcP) that correctly identified the tonB-dT protein in Avibacterium paragallinarum samples; R3) In Avibacterium paragallinarum: With the 2 peptides 4 clones of hybridomas producing monoclonal antibodies (Am) were obtained; however, only one clone (1G7G8) was reactive to tonB-dT. R4) With Am-1G7G8, a qualitative ICT prototype (presence and absence) was developed that proved to be specific for Avibacterium paragallinarum against aMPV samples and other avian pathogens. A TIC prototype was developed for the detection of Avibacterium paragallinarum without presenting cross-reactivity against aMPV and provides results within 30 minutes of applying the sample.

PE2018001160A 2018-06-19 2018-06-19 DIFFERENTIAL AND RAPID DIAGNOSTIC KIT FOR DETECTION OF INFECTIOUS CORIZA AND SWOLL HEAD SYNDROME IN THE POULTRY INDUSTRY PE20191840A1 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
PE2018001160A PE20191840A1 (en) 2018-06-19 2018-06-19 DIFFERENTIAL AND RAPID DIAGNOSTIC KIT FOR DETECTION OF INFECTIOUS CORIZA AND SWOLL HEAD SYNDROME IN THE POULTRY INDUSTRY
PCT/PE2019/000006 WO2019245392A1 (en) 2018-06-19 2019-06-13 Rapid differential diagnosis kit for detecting infectious coryza and swollen head syndrome in the poultry farming industry
ARP190101668A AR119649A1 (en) 2018-06-19 2019-06-18 DIFFERENTIAL AND RAPID DIAGNOSTIC KIT FOR THE DETECTION OF INFECTIOUS CORYZA AND SLOTHED HEAD SYNDROME IN THE POULTRY INDUSTRY

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PE2018001160A PE20191840A1 (en) 2018-06-19 2018-06-19 DIFFERENTIAL AND RAPID DIAGNOSTIC KIT FOR DETECTION OF INFECTIOUS CORIZA AND SWOLL HEAD SYNDROME IN THE POULTRY INDUSTRY

Publications (1)

Publication Number Publication Date
PE20191840A1 true PE20191840A1 (en) 2019-12-31

Family

ID=68983491

Family Applications (1)

Application Number Title Priority Date Filing Date
PE2018001160A PE20191840A1 (en) 2018-06-19 2018-06-19 DIFFERENTIAL AND RAPID DIAGNOSTIC KIT FOR DETECTION OF INFECTIOUS CORIZA AND SWOLL HEAD SYNDROME IN THE POULTRY INDUSTRY

Country Status (3)

Country Link
AR (1) AR119649A1 (en)
PE (1) PE20191840A1 (en)
WO (1) WO2019245392A1 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115029456B (en) * 2021-09-23 2023-04-07 北京市农林科学院 Transverse flow detection reagent and method for rapidly and sensitively detecting avibacterium paragallinarum nucleic acid

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102007414B (en) * 2008-02-08 2015-06-17 一般财团法人化学及血清疗法研究所 Method and kit for detection of anti-avibacterium paragallinarum antibody

Also Published As

Publication number Publication date
AR119649A1 (en) 2022-01-05
WO2019245392A1 (en) 2019-12-26

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