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BackgroundMyosin-binding protein C3 A31P (MYBPC3-A31P) missense mutation is a genetic deviation associated with the development of hypertrophic cardiomyopathy (HCM) in Maine Coon cats. The standard detection of the MYBPC3-A31P mutation is complicated, time-consuming, and expensive. Currently, there has been a focus on the speed and reliability of diagnostic tools. Therefore, this study aimed to develop a loop-mediated isothermal amplification assay (LAMP) coupled with a lateral flow dipstick (LFD) test to detect MYBPC3-A31P mutations in Maine Coon cats.Materials and MethodsFifty-five Maine Coon cats were enrolled in this study, and blood samples were collected. MYBPC3-A31P was genotyped by DNA sequencing. Primers for LAMP with a LFD test were designed. The optimal conditions were determined, including temperature and time to completion for the reaction. The sensitivity of A31P-LAMP detection was compared between agarose gel electrophoresis (the standard method) and the LFD test. The A31P-LAMP-LFD test was randomly performed on seven cats (four with the A31P mutation and three wild-type cats).ResultsThe A31P-LAMP procedure was able to distinguish between cats with MYBPC3-A31P wild-type cats and MYBPC3-A31P mutant cats. The LAMP reactions were able to be completed in 60 min at a single temperature of 64◦C. Moreover, this study demonstrated that A31P-LAMP coupled with the LFD test allowed for A31P genotype detection at a lower DNA concentration than agarose gel electrophoresis.DiscussionsThis new A31P-LAMP with a LFD test is a successful and reliable assay with a rapid method, cost-effectiveness, and low requirements for sophisticated equipment for the detection of MYBPC3-A31P mutations. Thus, this assay has excellent potential and can be recognized as a novel screening test for hypertrophic cardiomyopathy associated with MYBPC3-A31P mutations in felines.

(1) Background: The aim of this study was to find the prevalence of feline hip dysplasia (HD), patellar luxation and lumbosacral transitional vertebra (LTV) in pedigree cats in the Czech Republic. (2) Methods: 107 pedigree cats at least 10 months old were recruited prospectively at the Small Animal Clinic at the University of Veterinary and Pharmaceutical Sciences Brno, CZ, between April 2019 and July 2020. (3) Results: The prevalence of hip joint dysplasia in all pedigree cats was 46.7%, of which 78% of cats had bilateral dysplasia. The HD was mainly from mild (grade 1) to moderate (grade 2); however, 6.1% of hip joints showed signs of severe HD (grade 3) in Maine Coon and Siberian cats. Patellar luxation was noted in 32.7% of the pedigree cats, was present bilaterally in 91.4% and was grade 1 or 2 in most cats. The presence of LTV was noted in 7.5% of pedigree cats. (4) Conclusions: The high prevalence of HD in pedigree cats should be considered and screening pedigree cats for HD is recommended before they are used in breeding programs.

Background: Myosin-binding protein C3 A31P ( MYBPC3-A31P ) missense mutation is a genetic deviation associated with the development of hypertrophic cardiomyopathy (HCM) in Maine Coon cats. The standard detection of the MYBPC3-A31P mutation is complicated, time-consuming, and expensive. Currently, there has been a focus on the speed and reliability of diagnostic tools. Therefore, this study aimed to develop a loop-mediated isothermal amplification assay (LAMP) coupled with a lateral flow dipstick (LFD) test to detect MYBPC3-A31P mutations in Maine Coon cats., Materials and Methods: Fifty-five Maine Coon cats were enrolled in this study, and blood samples were collected. MYBPC3-A31P was genotyped by DNA sequencing. Primers for LAMP with a LFD test were designed. The optimal conditions were determined, including temperature and time to completion for the reaction. The sensitivity of A31P -LAMP detection was compared between agarose gel electrophoresis (the standard method) and the LFD test. The A31P -LAMP-LFD test was randomly performed on seven cats (four with the A31P mutation and three wild-type cats)., Results: The A31P -LAMP procedure was able to distinguish between cats with MYBPC3-A31P wild-type cats and MYBPC3-A31P mutant cats. The LAMP reactions were able to be completed in 60 min at a single temperature of 64◦C. Moreover, this study demonstrated that A31P -LAMP coupled with the LFD test allowed for A31P genotype detection at a lower DNA concentration than agarose gel electrophoresis., Discussions: This new A31P -LAMP with a LFD test is a successful and reliable assay with a rapid method, cost-effectiveness, and low requirements for sophisticated equipment for the detection of MYBPC3-A31P mutations. Thus, this assay has excellent potential and can be recognized as a novel screening test for hypertrophic cardiomyopathy associated with MYBPC3-A31P mutations in felines., Competing Interests: The research was conducted with a patent application (ID: 2103003416). The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Sukumolanan, Demeekul and Petchdee.)

Simple Summary: Hip dysplasia, patellar luxation and lumbosacral transitional vertebra are not well described in cats, most likely because cats can often much better compensate for pelvic limb lameness and hide their pain and, as a result, owners are less likely to notice this condition. Pedigree cats at least 10 months old were recruited prospectively in this study to find the prevalence of feline hip dysplasia, patellar luxation and lumbosacral transitional vertebra. The prevalence of hip joint dysplasia in all pedigree cats was 46.7%, of which 78% of cats had bilateral dysplasia. Dysplasia was mainly from mild (grade 1) to moderate (grade 2); however, 6.1% of hip joints showed signs of severe hip dysplasia (grade 3) in Maine Coon and Siberian cats. Patellar luxation was noted in 32.7% of the pedigree cats, was present bilaterally in 91.4% and was grade 1 or 2 in most cats. The presence of lumbosacral transitional vertebra was noted in 7.5% of the pedigree cats. The high prevalence of hip dysplasia in pedigree cats should be considered and screening pedigree cats for hip dysplasia is recommended before they are used in breeding programs. (1) Background: The aim of this study was to find the prevalence of feline hip dysplasia (HD), patellar luxation and lumbosacral transitional vertebra (LTV) in pedigree cats in the Czech Republic. (2) Methods: 107 pedigree cats at least 10 months old were recruited prospectively at the Small Animal Clinic at the University of Veterinary and Pharmaceutical Sciences Brno, CZ, between April 2019 and July 2020. (3) Results: The prevalence of hip joint dysplasia in all pedigree cats was 46.7%, of which 78% of cats had bilateral dysplasia. The HD was mainly from mild (grade 1) to moderate (grade 2); however, 6.1% of hip joints showed signs of severe HD (grade 3) in Maine Coon and Siberian cats. Patellar luxation was noted in 32.7% of the pedigree cats, was present bilaterally in 91.4% and was grade 1 or 2 in most cats. The presence of LTV was noted in 7.5% of pedigree cats. (4) Conclusions: The high prevalence of HD in pedigree cats should be considered and screening pedigree cats for HD is recommended before they are used in breeding programs. [ABSTRACT FROM AUTHOR]