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.
Highlights
Dengue is the most important arthropod-borne viral infection of humans with a large global burden
Detection rate of Simplexa Dengue compared to conventional reversetranscription and polymerase chain reaction (RT-PCR) and SYBR Green real-time reverse transcriptase (RT)-PCR
dengue virus (DENV) genome detections were performed in all samples
Summary
Dengue is the most important arthropod-borne viral infection of humans with a large global burden. There are an estimated 50 million infections per year occurring across approximately 100 countries in tropical and sub-tropical regions in the world with potential for wider distribution. The disease affects approximately 2.5 billion people living in Southeast Asia, the Pacific, and the Americas [1,2]. Dengue disease is caused by dengue virus (DENV), a member of Flaviviridae family, with a substantial genetic diversity shown by the presence of four serotypes (DENV-1, -2, -3, and -4) and multiple genotypes (or subtypes) within each serotype [4,5]. DENV is transmitted through a human-mosquito cycle with the aid of Aedes aegypti and Ae. albopictus mosquito vectors. The genome consists of single-stranded positive-sense RNA which encodes three structural (C, prM/M, E) and seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5) [1]
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