versão impressa ISSN 1813-5363
CAMPERO M., Marisol; BUSTAMANTE G., Zulema; VERAMENDI T., Silene e ROJAS B., Jorge. Development of a commercial molecular diagnosis for disease in chickens(Gallus gallus). BIOFARBO [online]. 2011, vol.19, n.1, pp. 34-43. ISSN 1813-5363.
Poultry breeding is one of the most widespread activity in the department of Cochabamba- Bolivia, where producers are permanently facing major challenges as the emergence of new diseases like mycoplasmosis caused by Mycoplasma gallisepticum and Mycoplasma synoviae. Mycoplamosis causes significant economic losses because this disease spreads very fast. Today the diagnosis of these pathogens is performed by serological methods; however, these techniques are time consuming and sometimes lead to misidentifications. Molecular techniques based on real-time PCR are more advantageous than serological techniques as they are rapid, sensitive and specific. Moreover they can be less expensive when analyzing various pathogens in a single assay (multiplex PCR). The aim of this study was to compare the different methods for molecular diagnosis of M. gallisepticum and M. synoviae to establish a commercial service. To achieve this goal, we used tracheal swab samples obtained from the airways. DNA was extracted by three methods (kit DNA extraction Mini signature Qiamp Qiagen, heat shock method and conventional method of lysis buffer) and then the samples were analyzed by a commercial kit (LSI TaqVet) and by a molecular diagnostic kit developed in this study. The method of extraction with a lysis buffer together with the molecular diagnostic kit developed in this study generated the best results and demonstrated better specificity, sensitivity and low cost for the molecular diagnosis of avian mycoplasmosis.
Palavras-chave : Real-time PCR; Taqman; SyberGreen; diagnostic; Mycoplasma.