A convenient and efficient microsatellite-based assay for resolving parentages in dogs

Area of Science:

• Veterinary Genetics
• Molecular Biology
• Canine Forensics

Background:

• Accurate parentage determination is crucial for canine breeding programs, genetic research, and breed registration.
• Existing parentage testing methods can be time-consuming and costly.
• The need for a standardized, efficient, and reliable canine parentage analysis is evident.

Purpose of the Study:

• To develop and validate a practical and efficient parentage analysis test for dogs.
• To establish a robust assay utilizing multiplex polymerase chain reaction (PCR) technology.
• To assess the accuracy and reliability of the developed test across different breeds.

Main Methods:

• Utilized ten polymorphic microsatellite markers in three multiplex-PCR reactions.
• Standardized PCR cycling conditions for all multiplexes.
• Employed a multiple-dye detection system and pooled PCR products for electrophoresis.
• Validated assay repeatability across four different thermal cyclers.
• Typed 100 dogs from four distinct breeds to evaluate efficiency.

Main Results:

• Achieved easily interpretable results across multiple thermal cyclers, demonstrating assay repeatability.
• Demonstrated high efficiency with 100 dogs typed from four breeds.
• Attained high exclusion probabilities, ranging from 99.34% to 99.93%, assuming one known parent.

Conclusions:

• The developed parentage analysis test is practical, efficient, and reliable for canine applications.
• The multiplex PCR approach offers a standardized and repeatable method for dog DNA typing.
• The high exclusion probabilities confirm the test's effectiveness in accurate parentage determination.