European Journal of Biomedical and Pharmaceutical Sciences

DEVELOPMENT AND VALIDATION OF A QUANTITATIVE MULTIPLEX REAL TIME PCR ASSAY FOR IDENTIFICATION OF BACTERIAL PATHOGENS FROM RESPIRATORY SPECIMENS

Ghazala Nathu*, Nitalia Naeem, Adila Nathu, N. Kazmi and R. Chandra

ABSTRACT

Development of timely and accurate methods for diagnosing bacterial respiratory tract infections has been a challenge for the clinical microbiology laboratory. Currently available commercial respiratory panels do not identify some important bacterial pathogens responsible for lower respiratory tract infections (LRTI). Moreover, in many instances, an accurate diagnosis cannot be made due to the overlapping clinical features of bacterial and viral infections on presentation. Culture methods are currently the gold standard for identification of LRTI pathogens. Molecular detection methodologies can offer more sensitive, rapid and targeted identification of these respiratory bacterial pathogens. At the lab, we have validated a laboratory developed sample to answer multiplex PCR for rapid detection of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis on the Luminex ARIES platform. The assay was evaluated for its analytical performance characteristics and its utility for patient testing in a reference laboratory. In order to provide a comprehensive molecular testing menu for detection of bacterial pathogens, the Aries Group A Strep IVD Assay, a real time PCR assay for detection of Group A Streptococcus pyogenes was also evaluated for its performance characteristics.

Keywords: LRTI, PCR, SPC.