title: Performance of Simplexa dengue molecular assay compared to conventional and SYBR green RT-PCR for detection of dengue infection in Indonesia.
abstract: Diagnostic tests based on detection of dengue virus (DENV) genome are available with varying sensitivities and specificities. The Simplexa Dengue assay (Focus Diagnostics) is a newly developed real-time RT-PCR method designed to detect and serotype DENV simultaneously. To assess the performance of the Simplexa Dengue assay, we performed comparison with conventional RT-PCR and SYBR Green real-time RT-PCR on patients sera isolated from eight cities across Indonesia, a dengue endemic country. A total of 184 sera that were confirmed using NS1 and/or IgM and IgG ELISA were examined. Using conventional and SYBR Green real-time RT-PCR, we detected DENV in 53 (28.8%) and 81 (44.0%) out of 184 sera, respectively. When the Simplexa Dengue assay was employed, the detection rate was increased to 76.6% (141 out of 184 samples). When tested in 40 sera that were confirmed by virus isolation as the gold standard, the conventional RT-PCR yielded 95% sensitivity while the sensitivity of SYBR Green real-time RT-PCR and Simplexa Dengue assay reached 97.5% and 100%, respectively. The specificities of all methods were 100% when tested in 43 non-dengue illness and 20 healthy human samples. Altogether, our data showed the higher detection rate of Simplexa Dengue compared to conventional and SYBR Green real-time RT-PCR in field/surveillance setting. In conclusion, Simplexa Dengue offers rapid and accurate detection and typing of dengue infection and is suitable for both routine diagnostic and surveillance.
authors.affiliation: Eijkman Institute for Molecular Biology, Jakarta, Indonesia.
authors.forename: R Tedjo
authors.initials: RT
authors.lastname: Sasmono
authors.affiliation: Clinical Pathology Department, School of Medicine and Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia.
authors.forename: Aryati
authors.initials: A
authors.lastname: Aryati
authors.affiliation: Clinical Pathology Department, School of Medicine and Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia.
authors.forename: Puspa
authors.initials: P
authors.lastname: Wardhani
authors.affiliation: Eijkman Institute for Molecular Biology, Jakarta, Indonesia.
authors.forename: Benediktus
authors.initials: B
authors.lastname: Yohan
authors.affiliation: Eijkman Institute for Molecular Biology, Jakarta, Indonesia; Agency for the Assessment and Application of Technology, Jakarta, Indonesia.
authors.forename: Hidayat
authors.initials: H
authors.lastname: Trimarsanto
authors.affiliation: Health Polytechnic, Jambi Provincial Health Office, Kotabaru, Jambi, Indonesia and Graduate School in Medicine, Diponegoro University, Semarang, Indonesia.
authors.forename: Sukmal
authors.initials: S
authors.lastname: Fahri
authors.affiliation: Eijkman Institute for Molecular Biology, Jakarta, Indonesia.
authors.forename: Tri Y
authors.initials: TY
authors.lastname: Setianingsih
authors.affiliation: Eijkman Institute for Molecular Biology, Jakarta, Indonesia.
authors.forename: Febrina
authors.initials: F
authors.lastname: Meutiawati
mesh_terms: D003715:Dengue / Q000175:diagnosis* / Q000235:genetics; D003716:Dengue Virus / Q000235:genetics* / Q000302:isolation & purification; D016679:Genome, Viral; D006801:Humans; D007214:Indonesia; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
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