Optimization of a duplex PCR for detection of Mycoplasma gallisepticum and Mycoplasma synoviae in tissue samples from Gallus gallus

Abstract

The objective of the work was to optimize duplex PCR for the detection of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) in tracheal tissue samples in Gallus gallus in order to save time and economic resources in their diagnosis. The methodology consisted of carrying out an in silico evaluation of the ideal primers for their detection, the first tests such as primer concentration, sensitivity and specificity were carried out, followed by determining the prevalence in samples from different poultry houses at national level during a period of 4 years. Once the first tests were completed, the ideal primers (MG 3 and MS 2) were chosen for duplex PCR, achieving a sensitivity and specificity greater than 99%. This technique was put into practice and 669 samples of chicken tracheal tissue were analyzed, in which 45 samples were found to be positive for MG and/or MS. Finally, the positive samples were sequenced in Macrogen-Korea. The prevalence of these pathogens was found to be low in the samples analyzed, but the result is of great importance because of the possibility of spreading these pathogens in poultry houses and causing great losses in the poultry sector.