Development of a Multiplex PCR for Detection of Common Causative Agent of Respiratory Tract Infections Include Streptococcus Pneumonia, Staphylococcus Aureus, KlebsiellaPneumonia and Mycobacterium Tuberculosis

Pneumonia is caused by a range of microorganisms. The incidence of bacterial pneumonia can be divided into two categories: hospital-acquired pneumonia and community-acquired pneumonia. Community acquired Pneumonia is a related leading cause in different populations. Staphylococcus aureus and Klebsiella pneumonia are the most common cause of hospital-acquired pneumonia, and Streptococcus pneumonia and mycobacterium tuberculosis are Cause of community-acquired pneumonia. The aim of this was to investigate the common causative factors of respiratory tract infections include Streptococcus pneumonia, Staphylococcus aureus, Klebsiella pneumonia and mycobacterium tuberculosis using multiplex PCR was the main aim of this study. This study was experimental development. Multiplex PCR is a rapid, accurate molecular method for the simultaneous detection of two or more specific genes in a reaction.In this study, multiplex PCR method for simultaneous detection in the genome-specific sequences leading causes of pneumonia (Streptococcus pneumonia, mycobacterium tuberculosis, Staphylococcus aurous and Klebsiella pneumonia) was done. Specific genes that we chose in this study were: ply (pneumolysin) for Streptococcus pneumonia, nuc (thermonuclease) for the staphylococcus aureus, gyrA (gyrase A) for Klebsiella pneumoniae and pncA gene for Mycobacterium tuberculosis, also the 16s ribosomal gene was used as a positive control. After set-up procedure and observing specific bands, sequencing was confirmed the accuracy of observed bands.Our results showed that the detection of Streptococcus pneumoniae, Staphylococcus aureus, Klebsiellapneumoniae and Mycobacterium tuberculosis in a Multiplex PCR reaction is possible. In fact, this method can be used for direct and simultaneous detection of these agents in the respiratory specimens.