Authors: SUNG WOO KIM, JIN SEOK CHOI, NEELESH SHARMA, YEOUNG GYU KO, YOON JUNG DO, MIJEONG BYUN, HWAN HOO SEONG, SOO BONG PARK, DONG KEE JEONG
Abstract: The monomorphism of newly hatched chicks is a major hurdle for poultry farmers at the time of sexing. The traditional sexing techniques are inaccurate and time-consuming. Therefore, the present study aimed to develop a loop-mediated isothermal amplification (LAMP)-based sex determination method to obtain a higher separation accuracy rate as compared to conventional methods. LAMP is a simple, quick, and relatively inexpensive diagnostic tool. Genomic DNA was extracted from the blood samples of Leghorn and Korean native chicks for LAMP. Two sets of primer were designed, consisting of four isothermal amplification primers in each, to amplify the female sex-specific sequence (HUR0417) of the PKCIW gene. We found that the corresponding female samples showed significant amplification of the PKCIW gene, while this was not observed in the male samples. Moreover, this method produced results in less than 1 h. The present investigation suggests that the LAMP-based sexing method is a sensitive, accurate, and time-saving method in chicks at an early age compared to the conventional sexing methods.
Keywords: Chicken, Bst DNA polymerase, isothermal amplification primers, loop-mediated isothermal amplification, PKCIW, sex determination
Full Text: PDF