Real-time PCR detection chemistry》.pdf

Clinica Chimica Acta 439 (2015) 231–250 Contents lists available at ScienceDirect Clinica Chimica Acta journal homepage: www. else vi er. com/ locate /clinchim Invited critical review Real-time PCR detection chemistry E. Navarro a,⁎, G. Serrano-Heras a, M.J. Castaño a, J. Solera b a Research Unit, General University Hospital, Laurel s/n, 02006 Albacete, Spain b Internal Medicine Department, General University Hospital, Hermanos Falcó 37, 02006 Albacete, Spain a r t i c l e i n f o a b s t r a c t Article history: Real-time PCR is the method of choice in many laboratories for diagnostic and food applications. This technology Received 18 March 2014 merges the polymerase chain reaction chemistry with the use of fluorescent reporter molecules in order to Received in revised form 9 October 2014 monitor the production of amplification products during each cycle of the PCR reaction. Thus, the combination Accepted 11 October 2014 of excellent sensitivity and specificity, reproducible data, low contamination risk and reduced hand-on time, Available online 22 October 2014 which make it a post-PCR analysis unnecessary, has made real-time PCR technology an appealing alternative Keywords: to conventional PCR. The present paper attempts to provide a rigorous overview of fluorescent-base