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Bovine CLEC7A genetic variants and their association with seropositivity in Johne's disease ELISA
Authors:SD Pant  CP Verschoor  FS Schenkel  Q You  DF Kelton  NA Karrow
Institution:1. Centre for Genetic Improvement of Livestock, Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario N1G 2W1, Canada;2. Population Medicine, Ontario Veterinary College, University of Guelph, Guelph, Ontario N1G 2W1, Canada
Abstract:Mycobacterium avium ssp. paratuberculosis (MAP) infection in cattle causes significant economic losses to the dairy and beef industries resulting from reduced productivity, premature culling and mortality. Bovine Dectin-1, an important pattern recognition molecule that is able to generate a proinflammatory response by acting alongside Toll like receptor (TLR) signaling, is known to co-operate with TLR2 to specifically activate a macrophage proinflammatory response against mycobacterial infections. Therefore, the goal of this study was to identify single nucleotide polymorphisms (SNPs) in the gene encoding bovine Dectin-1 (CLEC7A) and to assess their association with susceptibility to MAP infection in dairy cattle. Blood and milk samples, collected from commercial dairy operations, were tested for MAP infection using blood and milk ELISAs and a resource population consisting of 197 infected and 242 healthy cattle was constructed. Pooled DNA was used for sequencing and eight single nucleotide polymorphisms (SNPs) were identified. Identified SNPs were genotyped on the resource population using the iPLEX MassARRAY system and statistical analysis was performed using logistic regression fitting the additive and dominance effects of each SNP in the model. Out of a total of eight identified SNPs, five were successfully genotyped, and three out of these five SNPs were found to be in complete linkage. Statistical analysis revealed a strong association between a non-synonymous SNP c.589A>G (p = 0.008), and MAP infection status of the resource population inferred by seropositivity in MAP antibody specific ELISAs. This SNP c.589A>G was located in the geneic region that encodes the carbohydrate recognition domain of bovine Dectin-1. Therefore, further investigation of its functional relevance is warranted.
Keywords:AIC  Akaike's information criterion  BTA  Bos taurus autosome  CD  Crohn's disease  CLEC7A  C-type lectin domain family 7 member A/Dectin-1  CTL  C-type lectin  D  Linkage disequilibrium  ELISA  Enzyme linked immuno sorbent assay  IFNG  Interferon-γ  IL12  Interleukin-12  ITAM  Immune receptor tyrosine-based activation motif  JD  Johne's disease  MAP  Mycobacterium avium ssp  paratuberculosis  NC  Negative control  OD  Optical density  OR  Odds ratio  PC  Positive control  PCR  Polymerase chain reaction  PRM  Pattern recognition molecule  QTL  Quantitative trait loci  S  Sample  SNP  Single nucleotide polymorphism  SP110  SP110 nuclear body protein  Th1  T-helper 1  TLR  Toll like receptor
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