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Background  The prevalence of gastrointestinal (GI) neoplastic polyps in Jack Russell terriers (JRTs) has increased in Japan since the late 2000s. Recently, we demonstrated that JRTs with GI polyps harbor identical germline variant in the  APC  gene (c.[462_463delinsTT]) in the heterozygous state. Thus, this disease is an autosomal dominant hereditary disorder. Although the affected JRTs have distinct features, such as the development of multiple GI polyps and an early age of disease onset, genetic testing is indispensable for a definitive diagnosis. Here, polymerase chain reaction (PCR)-based assays capable of detecting germline  APC  variant were designed and validated using synthetic wild-type and mutant DNAs and genomic DNAs from carrier and non-carrier dogs.    Result  First, the PCR-restriction fragment length polymorphism (PCR-RFLP) assay was developed by taking advantage of the germline  APC  variant creating a new restriction site for  Mse I. In the PCR-RFLP assay, the 156-bp region containing the variant site was amplified by PCR and subsequently digested with  Mse I, yielding diagnostic 51 and 58 bp fragments from the mutant allele and allowing determination of the  APC  genotypes. It was possible to determine the genotypes using genomic DNA extracted from the peripheral blood, buccal swab, or formalin-fixed paraffin-embedded tissue. Next, a TaqMan duplex real-time PCR assay was developed, where a 78-bp region flanking the variant was amplified in the presence of wild-type allele- and mutant allele-specific fluorescent probes. Using blood-derived DNA, altogether 40 cycles of PCR amplification determined the  APC  genotypes of all examined samples by measuring the fluorescence intensities. Importantly, false-positive and false-negative errors were never detected in both assays.    Conclusion  In this study, we developed highly reliable genetic tests for hereditary GI polyposis in JRTs, providing accurate assessment of the presence of the causative germline  APC  variant. The genotyping assays could contribute to the diagnosis and prevention of hereditary GI polyposis in dogs.    Supplementary Information  The online version contains supplementary material available at 10.1186/s12917-020-02731-7.

PCR-based genotyping assays to detect germline APC variant associated with hereditary gastrointestinal polyposis in Jack Russell terriers