Development of a multiplex allele-specific PCR assay for rapid screening of Lactoferrin TATA box polymorphisms

Lactoferrin, a natural antimicrobial compound found in a variety of mucosal surface, is a potential candidate gene in cattle for increasing resistance to various infectious diseases. Previously reported A/C polymorphism (-28 A/C) within TATA box sequence of lactoferrin (LTF) gene was shown to be a key factor in lactoferrin expression level. The objective of this study was to develop an allele-specific PCR method for high-throughput screening of single nucleotide polymorphisms at lactoferrin TATA box in dairy cattle. The technique involves PCR amplification of specific alleles that differ only in one base. This approach, which was validated on three known TATA box genotypes, is accurate, sensitive, and suitable for high-throughput screening since it requires only one reaction per case; it is performed in a single tube and also it needs minimal sample processing. © 2012 Springer-Verlag London Limited.