title: Comparison of an in-house real-time RT-PCR assay with a commercial assay for detection of enterovirus RNA in clinical samples.
abstract: Molecular detection of enterovirus (EV) RNA based on PCR methods is a quicker and more sensitive approach than culture methods. At present, different PCR-based methods for EV RNA detection are available, but comparisons of results obtained according to the different approaches are limited. We evaluated an in-house real-time RT-PCR assay with a commercialized TaqMan real-time RT-PCR kit for detection of EV. Consecutive clinical specimens from paediatric patients less than 6 years old with clinical suspicion of EV infection were analyzed between July and November 2010. After RNA extraction, samples were amplified both by the real-time RT-PCR commercial assay and the in-house assay. A total of 19 of 132 patients (14.4%) involving 20 samples (14 plasma samples and 6 CSF) were positive in at least one of the two assays. The sensitivity of the in-house assay when the MutaPLATE® assay was used as a reference was 90% (IC 95%; 74.35-100) and the specificity was 100% (IC 95%; 99.63-100). Cts results of two methods were statistically correlated (r = 0.774; P = 0.01). In conclusion, these two real-time RT-PCR assays are rapid and easy methods for detection of EV.
authors.affiliation: Department of Molecular Microbiology, Hospital Universitari Sant Joan de Deu, Esplugues, Barcelona, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Selva
authors.forename: A
authors.initials: A
authors.lastname: Martinez-Planas
authors.forename: J-J
authors.initials: JJ
authors.lastname: García-García
authors.forename: R
authors.initials: R
authors.lastname: Casadevall
authors.forename: C
authors.initials: C
authors.lastname: Luaces
authors.forename: C
authors.initials: C
authors.lastname: Muñoz-Almagro
mesh_terms: D002555:Cerebrospinal Fluid / Q000821:virology; D002648:Child; D002675:Child, Preschool; D019411:Clinical Laboratory Techniques / Q000379:methods*; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000175:diagnosis*; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D010949:Plasma / Q000821:virology; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods
chemical_list: D012367:RNA, Viral