American Association for Science and Technology AASCIT Communications Volume 1, Issue 3 October 20, 2014 online Rapid Diagnosis by Multiplex PCR Technology at Patients with Acute Respiratory Infections Aurelian Udristioiu, Manole Cojocaru Keywords: Antibiotic Resistance, PCR, Gene, AND, Resistance Markers, Microbiology Report ntroduction: PCR offer a number of potential advantages, results are available in a matter of hours rather than days, the extreme sensibility facilitates detection of even minutes the amounts of pathogen DNA in clinical samples and the test is not significantly affected by prior administration of antibiotics. Aim: The aim of this work was to rapidly identify the antibiotic resistance the monitoring of pathogen growth at the patients admitted in Hospitalization Intensive Care Unit of Emergency County Hospital Targu Jiu starting in December/2013. Method: The Analyzer Unyvero™ Pneumonia Application was used in detection of pneumonia associated pathogens and their antibiotic resistance genes using the Unyvero™ System following PCR, pathogen species with sequencing of the amplified microbial DNA. Pneumonia Cases Report December/2013: –1. Gender male, age 72 years, Sample ID 1311_1, the results was positive for Streptococcus pneumonia(873), Moraxela cataralis (308), Resistance markers of the Unyvero™ Pneumonia Panel: -tem β- Lactams gram-negative bacteria, -ermB Makrolides / Lincosamides, mecA Oxacillin Staphylococcus. Therapeutic failure must be considered when is administering Makrolides / Lincosamides(ermB), Macrolides (mefA), Penicilins (tem), [Figure 1, 2, 3]. -2. Gender male, age 65 years, Sample ID 1310_1, the results was positive for Klebsiella pneumonia(62), resistance marker result ermB ermB Makrolides / Lincosamides. Therapeutic failure must be considered when is administering Penicilins. -3. Gender Female age 55 years, Sample ID 1310_2, the results was positive Staphylococcus Aureus, tem β-Lactams gram-negative bacteria, -ermB Makrolides. Therapeutic failure must be considered when is administering Penicilins. Conclusions: The Unyvero™ results were available 2 days before the primary microbiology report and 3 days before the final confirmation results, obtained by microbiology culture. The Unyvero Analyzer only provides rapid data to support the therapeutic decision of currant medic. Introduction While microbiological culture is likely to remain a gold standard for infection diagnosis, there is growing interest at the potential of PCR technology to provide early, time critical information based on detection and recognition of bacterial or fungal pathogen DNA. Following PCR, pathogen species present can be identified sequencing of the amplified DNA. The assay for detection and identification of a defined panel of 25 bacterial of 25 bacterial and fungal pathogen known to cause a majority of acute or chronic respiratory infections. The need of sophisticated arsenal of new technological tools in the microbiology lab is dictated by two recent events. The first is the emergence and growth of though strain of microbe resistant to antibiotics, even the new antibiotics is drying up (ex. Microorganism such as K. pneumonia and E. coli have become resistant to third generation cephalosporins as well as carbapenems. The focus of researches now is to move beyond detecting single analytes to multiplex targets and detect more pathogens from a single specimen, ex. Sputum, (1). This application is a semi- quantitative nucleic acid test on the basis of eight multiplex PCR reactions that are performed in parallel. It is tested for the simultaneous detection and identification of multiple pathogen- derived nucleic acids in sputa, respiratory aspirates and bronchiallavage from individuals suspected of pneumonia, in order to provide information on pathogen species and antibiotic resistance genes. DNA from individuals exhibiting signs and symptoms of pneumonia aids in the diagnosis of respiratory bacterial or fungal infections if used in conjunction with other clinical and laboratory findings. Concomitant cultures are necessary to recover targeted organisms for further susceptibility testing. It is recommended that specimens found to be negative for pathogens and resistance genes after examination using the Unyvero™ Pneumonia Application must be confirmed by microbiological culture, [Photo 1]. Positive results do not rule out fungal or viral co-infection, or co-infection with other bacteria not present on the Unyvero™ Pneumonia panel and the agent detected may not be the definite cause of disease, (2). I