Development and Validation of KASP Assays for the Genotyping of Racing Performance-Associated Single Nucleotide Polymorphisms in Pigeons

Ching-Chi Chang; Benji Brayan I Silva; Huai-Ying Huang; Ching-Yi Tsai; Ronilo Jose D Flores; Lemmuel L Tayo; Yu-Chang Tyan; Ming-An Tsai; Gail Everette M Catulin; Kuo-Pin Chuang; Jenq-Lin Yang

doi:10.3390/genes12091383

Development and Validation of KASP Assays for the Genotyping of Racing Performance-Associated Single Nucleotide Polymorphisms in Pigeons

Genes (Basel). 2021 Sep 3;12(9):1383. doi: 10.3390/genes12091383.

Authors

Ching-Chi Chang¹, Benji Brayan I Silva², Huai-Ying Huang^{1

3}, Ching-Yi Tsai^{1

2}, Ronilo Jose D Flores^{4

5}, Lemmuel L Tayo⁶, Yu-Chang Tyan^{2

7

8

9

10

11}, Ming-An Tsai^{12

13}, Gail Everette M Catulin⁶, Kuo-Pin Chuang^{1

2

14

15}, Jenq-Lin Yang¹⁶

Affiliations

¹ Graduate Institute of Animal Vaccine Technology, College of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung 912, Taiwan.
² International Degree Program in Animal Vaccine Technology, International College, National Pingtung University of Science and Technology, Pingtung 912, Taiwan.
³ Demin Veterinary Hospital, Kaohsiung 807, Taiwan.
⁴ Institute of Biological Sciences, College of Arts and Sciences, University of the Philippines Los Baños, Laguna 4031, Philippines.
⁵ Graduate School, University of the Philippines Los Baños, Laguna 4031, Philippines.
⁶ School of Chemical, Biological and Materials Engineering and Sciences, Mapúa University, Intramuros, Manila 1002, Philippines.
⁷ School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
⁸ Institute of Medical Science and Technology, National Sun Yat-Sen University, Kaohsiung 804, Taiwan.
⁹ Department of Medical Imaging and Radiological Sciences, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
¹⁰ Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan.
¹¹ Research Center for Environmental Research, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
¹² Department of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung 912, Taiwan.
¹³ International Program in Ornamental Fish Technology and Aquatic Animal Health, International College, National Pingtung University of Science and Technology, Pingtung 912, Taiwan.
¹⁴ Research Center for Animal Biologics, National Pingtung University of Science and Technology, Pingtung 912, Taiwan.
¹⁵ School of Dentistry, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
¹⁶ Institute for Translation Research in Biomedicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung 807, Taiwan.

Abstract

Pigeon racing's recent upturn in popularity can be attributed in part to the huge prize money involved in these competitions. As such, methods to select pigeons with desirable genetic characteristics for racing or for selective breeding have also been gaining more interest. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for genotyping-specific genes is one of the most commonly used molecular techniques, which can be costly, laborious and time consuming. The present study reports the development of an alternative genotyping method that employs Kompetitive Allele Specific Polymerase Chain Reaction (KASP) technology with specifically designed primers to detect previously reported racing performance-associated polymorphisms within the LDHA, MTYCB, and DRD4 genes. To validate, KASP assays and PCR-RFLP assays results from 107 samples genotyped for each of the genes were compared and the results showed perfect (100%) agreement of both methods. The developed KASP assays present an alternative rapid, reliable, and cost-effective method to identify polymorphisms in pigeons.

Keywords: DRD4 gene; KASP; LDHA gene; MTCYB gene; PCR-RFLP; genetic resources; polymorphism.

Publication types

Research Support, Non-U.S. Gov't
Validation Study

MeSH terms

Animals
Columbidae / genetics
Columbidae / physiology*
Flight, Animal / physiology*
Genotyping Techniques / methods*
L-Lactate Dehydrogenase / genetics
Polymerase Chain Reaction / methods
Polymorphism, Restriction Fragment Length
Polymorphism, Single Nucleotide*
Receptors, Dopamine D4 / genetics

Substances

Receptors, Dopamine D4
L-Lactate Dehydrogenase