Eclipse probe assay for the detection of mycoplasma pneumoniae.

Mycoplasma pneumoniae is an important human pathogen that causes community acquired pneumonias and upper respiratory tract infections. Traditional detection techniques, such as culture and serology, cannot rapidly aid in the diagnosis of an acute infection with adequate sensitivity and specificity. PCR testing has therefore become a popular testing method. However, lack of a standardized assay that is both rapid and costeffective has discouraged routine clinical testing, leading to misdiagnosis and inappropriate antibiotic therapy. Here we detail a real-time PCR assay for M. pneumoniae detection that utilizes an Eclipse hybridization probe (Nanogen) and targets the P1 cytadeherence gene. We enhanced turnaround time and cost-efficiency by developing a high-throughput protocol that extracts all respiratory sample types simultaneously.