MarÃa Karla MartÃnez
Faculty of Biology. University of Havana. Cuba
Title: EVALUATION AND INTRODUCTION OF REAL TIME PCR NEW ASSAYS FOR THE DIAGNOSTICAND SURVEILLANCE OF HUMAN RESPIRATORY VIRUS
Biography
Biography: MarÃa Karla MartÃnez
Abstract
This study proposed theevaluation of four multiplex real timePCR for the diagnosis of 15 human respiratory viruses causing acute respiratory infection, as well as the introduction of the optimized system for the diagnosis and laboratory surveillance. For the optimization of multiplex real time PCR 352 nasopharyngeal swabs from July-August 2013 were used, multiplex RT-PCR was used as ¨gold standard¨ technique. Sensitivity, specificity, positive and negative predictive values and kappa index of multiplex real time PCR was determined. Efficiency of simple and multiplex real time PCR was calculated by calibration curve and slope determination. In addition, laboratory diagnosis in the period September 2013 to May 2014 with the optimized multiplex real time PCR was performed. Of the total of samples processed for optimization, 162 were positive by multiplex real time PCR and 112 by gold standard technique. Sensitivity, specificity, positive predictive value, negative predictive value and average kappa of the evaluation assays was 100%, 98.4%, 67%, 100% and 0.8, respectively. Furthermore, efficiency values for multiplex real time PCR systems were in the range 90.30 % to 103.09 % similar to those obtained by the simple system. Introduction of optimized assays allowed the detection of 1290 clinical samples positive for respiratory viruses, with the highest positivity percentage for human respiratory syncytial virus, 47.83%. In summary, multiplex real time PCR was more sensitive than multiplex RT-PCR and the efficiency values were similar to the simple real time PCR. These optimized systems allowed to update the algorithm for the diagnosis and surveillance of respiratory viruses in Cuba.