Detection of the Brachyspina mutation in Uruguayan Holstein cows using real time PCR and melting curve analysis

Detecção da mutação da Brachyspina em vacas Holandês Uruguaias usando PCR em tempo real e análise da curva de fusão

ABSTRACT:

Brachyspina syndrome (BS) is a rare monogenic autosomal recessive hereditary disorder of the Holstein Fresian breed caused by a deletion of 3.3Kb in the Fanconi anemia complementation group I (FANCI) gene on BTA-21, which leads to a frame-shift and premature stop codon. Some of the consequences of BS are the reduction of the fertility rate and milk production. This study developed a simple, sensitive, rapid cost- effective assay method based on real time PCR and melting curve analysis for the detection of BS carrier animals. Sixty-eight normal homozygous and four heterozygous carrier genotypes were detected and confirmed through traditional PCR- electrophoresis analysis. We concluded that the assay we have developed proved to be a reliable, highly precise and low-cost tool, which could be used to monitor the presence of the BS mutation in uruguayan Holstein breed.

Key words:
Bos taurus; Holstein; Brachyspina syndrome; real time PCR; melting curve analysis