<?xml version="1.0" encoding="UTF-8"?><xml><records><record><source-app name="Biblio" version="7.x">Drupal-Biblio</source-app><ref-type>17</ref-type><contributors><authors><author><style face="normal" font="default" size="100%">Goris, Nesya</style></author><author><style face="normal" font="default" size="100%">Frank Vandenbussche</style></author><author><style face="normal" font="default" size="100%">Herr, Cécile</style></author><author><style face="normal" font="default" size="100%">Villers, Jérôme</style></author><author><style face="normal" font="default" size="100%">Yves Van der Stede</style></author><author><style face="normal" font="default" size="100%">Kris De Clercq</style></author></authors></contributors><titles><title><style face="normal" font="default" size="100%">Validation of two real-time RT-PCR methods for foot-and-mouth disease diagnosis: RNA-extraction, matrix effect, uncertainty of measurement and precision.</style></title><secondary-title><style face="normal" font="default" size="100%">J Virol Methods</style></secondary-title><alt-title><style face="normal" font="default" size="100%">J. Virol. Methods</style></alt-title></titles><keywords><keyword><style  face="normal" font="default" size="100%">Animals</style></keyword><keyword><style  face="normal" font="default" size="100%">blood</style></keyword><keyword><style  face="normal" font="default" size="100%">Diagnostic Errors</style></keyword><keyword><style  face="normal" font="default" size="100%">Foot</style></keyword><keyword><style  face="normal" font="default" size="100%">Foot-and-Mouth Disease</style></keyword><keyword><style  face="normal" font="default" size="100%">Foot-and-Mouth Disease Virus</style></keyword><keyword><style  face="normal" font="default" size="100%">Reverse Transcriptase Polymerase Chain Reaction</style></keyword><keyword><style  face="normal" font="default" size="100%">RNA, Viral</style></keyword><keyword><style  face="normal" font="default" size="100%">Sensitivity and Specificity</style></keyword></keywords><dates><year><style  face="normal" font="default" size="100%">2009</style></year><pub-dates><date><style  face="normal" font="default" size="100%">2009 Sep</style></date></pub-dates></dates><volume><style face="normal" font="default" size="100%">160</style></volume><pages><style face="normal" font="default" size="100%">157-62</style></pages><language><style face="normal" font="default" size="100%">eng</style></language><abstract><style face="normal" font="default" size="100%">&lt;p&gt;Real-time reverse transcription polymerase chain reaction (rRT-PCR) assays are being used routinely for diagnosing foot-and-mouth disease virus (FMDV). Although most laboratories determine analytical and diagnostic sensitivity and specificity, a thorough validation in terms of establishing optimal RNA-extraction conditions, matrix effect, uncertainty of measurement and precision is not performed or reported generally. In this study, different RNA-extraction procedures were compared for two FMDV rRT-PCRs. The NucleoSpin columns available commercially combined high extraction efficiency with ease-of-automation. Furthermore, six different FMDV-negative matrices were spiked with a dilution series of FMDV SAT1 ZIM 25/89. Compared to cell-culture-spiked viral control samples, no matrix effect on the analytical sensitivity was found for blood or foot epithelium. Approximately 1log(10) reduction in detection limit was noted for faecal and tongue epithelium samples, whereas a 3log(10) decrease was observed for spleen samples. By testing the same dilution series in duplicate on 10 different occasions, an estimation of uncertainty of measurement and precision was obtained using blood as matrix. Both rRT-PCRs produced highly precise results emphasising their potential to replace conventional virological methods. The uncertainty measurement, as described in this study, proved to be a useful tool to evaluate the probability of making a wrong decision.&lt;/p&gt;</style></abstract><issue><style face="normal" font="default" size="100%">1-2</style></issue><custom1><style face="normal" font="default" size="100%">http://www.ncbi.nlm.nih.gov/pubmed/19447138?dopt=Abstract</style></custom1></record></records></xml>