Use of recombinant S1 spike polypeptide to develop a TCoV-specific antibody ELISA

Maged H. Gomaa; Dongwan Yoo; Davor Ojkic; John R. Barta

doi:10.1016/j.vetmic.2009.04.010

Use of recombinant S1 spike polypeptide to develop a TCoV-specific antibody ELISA

Maged H. Gomaa, Dongwan Yoo, Davor Ojkic, John R. Barta

Research output: Contribution to journal › Article › peer-review

Abstract

Turkey coronavirus (TCoV) causes diarrhoea in young turkey poults but little is known about its prevalence in the field. To address this, a portion of the S1 region of the spike glycoprotein of TCoV carrying relevant B cell epitopes (amino acid positions 54-395) was cloned and expressed in Escherichia coli. This protein was purified and used to develop an indirect ELISA for detection of antibodies against TCoV. Using experimentally derived positive and negative turkey serum samples this ELISA showed high sensitivity (95%) and specificity (92%) for TCoV. To further evaluate the potential of the ELISA, 360 serum samples from commercial turkey farms in Ontario were tested for TCoV-specific antibodies using the recombinant TCoV ELISA. High seroprevalence of TCoV was found with 71.11% of breeders and 56.67% of meat turkeys testing seropositive. Although there was significant positive correlation with a TCoV-N protein-based ELISA, there was little to no correlation with the whole IBV antigen-based ELISA when field sera were tested for antibodies against TCoV.

Original language	English (US)
Pages (from-to)	281-288
Number of pages	8
Journal	Veterinary Microbiology
Volume	138
Issue number	3-4
DOIs	https://doi.org/10.1016/j.vetmic.2009.04.010
State	Published - Sep 18 2009

Keywords

Antibody
Enzyme immunoassay
IBV
Seroprevalence
Spike protein
TCoV

ASJC Scopus subject areas

Microbiology
veterinary(all)

Online availability

10.1016/j.vetmic.2009.04.010

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@article{277db87ae29d42c3b29b656c7f4da8e9,

title = "Use of recombinant S1 spike polypeptide to develop a TCoV-specific antibody ELISA",

abstract = "Turkey coronavirus (TCoV) causes diarrhoea in young turkey poults but little is known about its prevalence in the field. To address this, a portion of the S1 region of the spike glycoprotein of TCoV carrying relevant B cell epitopes (amino acid positions 54-395) was cloned and expressed in Escherichia coli. This protein was purified and used to develop an indirect ELISA for detection of antibodies against TCoV. Using experimentally derived positive and negative turkey serum samples this ELISA showed high sensitivity (95%) and specificity (92%) for TCoV. To further evaluate the potential of the ELISA, 360 serum samples from commercial turkey farms in Ontario were tested for TCoV-specific antibodies using the recombinant TCoV ELISA. High seroprevalence of TCoV was found with 71.11% of breeders and 56.67% of meat turkeys testing seropositive. Although there was significant positive correlation with a TCoV-N protein-based ELISA, there was little to no correlation with the whole IBV antigen-based ELISA when field sera were tested for antibodies against TCoV.",

keywords = "Antibody, Enzyme immunoassay, IBV, Seroprevalence, Spike protein, TCoV",

author = "Gomaa, {Maged H.} and Dongwan Yoo and Davor Ojkic and Barta, {John R.}",

note = "Funding Information: We thank Tony Cengija and Barb Mitchell of the Campus Animal Facility's Isolation Unit of the University of Guelph for the excellent care of our experimental animals and Stephanie Reabel and Inas Elawadli for assistance with animal handling and sample collection. We also thank Janet Swinton, Gaye Smith, Hongwen Fu, and Keith Harron of the University of Guelph's Animal Health Laboratory for technical assistance and Jeff Zhang from Cold Spring Farms for the field turkey sera. Hybrid Turkeys (Kitchener, ON, Canada) generously donated day of age poults used in the study. Dr. Billy Hargis, University of Arkansas, Fayetteville, AR, USA kindly provided SPF turkey serum samples. This work is supported by grants to JRB from the Natural Sciences and Engineering Council of Canada (NSERC) and the Ontario Ministry of Agriculture and Food and Rural Affairs (OMAFRA). MG is a scholarship recipient from the Egyptian Cultural and Educational Bureau of the Arab Republic of Egypt in Canada.",

year = "2009",

month = sep,

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doi = "10.1016/j.vetmic.2009.04.010",

language = "English (US)",

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AU - Yoo, Dongwan

AU - Ojkic, Davor

AU - Barta, John R.

N1 - Funding Information: We thank Tony Cengija and Barb Mitchell of the Campus Animal Facility's Isolation Unit of the University of Guelph for the excellent care of our experimental animals and Stephanie Reabel and Inas Elawadli for assistance with animal handling and sample collection. We also thank Janet Swinton, Gaye Smith, Hongwen Fu, and Keith Harron of the University of Guelph's Animal Health Laboratory for technical assistance and Jeff Zhang from Cold Spring Farms for the field turkey sera. Hybrid Turkeys (Kitchener, ON, Canada) generously donated day of age poults used in the study. Dr. Billy Hargis, University of Arkansas, Fayetteville, AR, USA kindly provided SPF turkey serum samples. This work is supported by grants to JRB from the Natural Sciences and Engineering Council of Canada (NSERC) and the Ontario Ministry of Agriculture and Food and Rural Affairs (OMAFRA). MG is a scholarship recipient from the Egyptian Cultural and Educational Bureau of the Arab Republic of Egypt in Canada.

PY - 2009/9/18

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N2 - Turkey coronavirus (TCoV) causes diarrhoea in young turkey poults but little is known about its prevalence in the field. To address this, a portion of the S1 region of the spike glycoprotein of TCoV carrying relevant B cell epitopes (amino acid positions 54-395) was cloned and expressed in Escherichia coli. This protein was purified and used to develop an indirect ELISA for detection of antibodies against TCoV. Using experimentally derived positive and negative turkey serum samples this ELISA showed high sensitivity (95%) and specificity (92%) for TCoV. To further evaluate the potential of the ELISA, 360 serum samples from commercial turkey farms in Ontario were tested for TCoV-specific antibodies using the recombinant TCoV ELISA. High seroprevalence of TCoV was found with 71.11% of breeders and 56.67% of meat turkeys testing seropositive. Although there was significant positive correlation with a TCoV-N protein-based ELISA, there was little to no correlation with the whole IBV antigen-based ELISA when field sera were tested for antibodies against TCoV.

AB - Turkey coronavirus (TCoV) causes diarrhoea in young turkey poults but little is known about its prevalence in the field. To address this, a portion of the S1 region of the spike glycoprotein of TCoV carrying relevant B cell epitopes (amino acid positions 54-395) was cloned and expressed in Escherichia coli. This protein was purified and used to develop an indirect ELISA for detection of antibodies against TCoV. Using experimentally derived positive and negative turkey serum samples this ELISA showed high sensitivity (95%) and specificity (92%) for TCoV. To further evaluate the potential of the ELISA, 360 serum samples from commercial turkey farms in Ontario were tested for TCoV-specific antibodies using the recombinant TCoV ELISA. High seroprevalence of TCoV was found with 71.11% of breeders and 56.67% of meat turkeys testing seropositive. Although there was significant positive correlation with a TCoV-N protein-based ELISA, there was little to no correlation with the whole IBV antigen-based ELISA when field sera were tested for antibodies against TCoV.

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KW - Enzyme immunoassay

KW - IBV

KW - Seroprevalence

KW - Spike protein

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JO - Veterinary Microbiology

JF - Veterinary Microbiology

IS - 3-4

ER -

Use of recombinant S1 spike polypeptide to develop a TCoV-specific antibody ELISA

Abstract

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