Development of two novel SYBR green-based quantitative PCR assays for detection andquantification of Mycoplasma gallisepticum and Mycoplasma synoviae

Mycoplasma gallisepticum (MG) and synoviae (MS) are considered the most pathogenic economically devastating avian mycoplasmas. A definite MG MS diagnosis depends on three main approaches, including isolation identification, demonstration of specific antibodies, detection genetic materials. fast precise technique is an indispensable prerequisite to prevent control pathogens more effectively. This study aimed develop validate convenient SYBR green-based quantitative PCR (qPCR) assays using genome primers rapidly detect quantify MS. Two sets were designed based highly conserved regions mgc2 vlhA genes in MS, respectively. The efficiencies both reactions calculated around 96% with a limit as few 10 copy numbers DNA/μL sample. standard curves showed linearity over dynamic range 6 log units from 10$^{1}$ 10$^{6}$ DNA numbers/μL without any significant variations inter- intra-assays. melting temperatures amplicons for determined be 79.90 °C 80.79 °C, results that only detected single melt peak cross reaction other non-targeted agents. developed further applied 60 swab specimens commercial poultry unknown statuses compared conventional assays. current 1 11 samples However, five positive tested gel-based protocols here have potential use diagnostic research tool clinical samples. Furthermore, proposed qPCR techniques can provide alternative approach surveillance epidemiological studies.