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.

Dengue viruses circulating in Indonesia: A systematic review and phylogenetic analysis of data from five decades

“…A full assessment of the 120 references resulted in the exclusion of 53 references due to the following: full texts were unavailable, and the available abstracts did not include adequate information to satisfy our outcome, the full text did not include adequate or complete serotype data, and nonstandard methods were used for serotyping. This resulted in 67 eligible studies, of which 13 references were from national journals . An additional 11 studies were included from reference list .…”

“…In total, 78 eligible studies were included for DENV serotype analysis . Of all the eligible studies, four studies were conducted based on mosquito isolates while the remaining studies assessed DENV isolates from human samples . Sixty two studies investigated dengue cases among local inhabitants of Indonesia,…”

“…Between 2008 and 2012, however, DENV‐1 become the dominant serotype in most Indonesian regions (Table ). Data of traveller‐associated dengue from Australia, Taiwan, Germany, and Romania support this finding.…”

Clinical and virological characteristics of dengue in Surabaya, Indonesia

“…DENV nucleic acid detection and serotyping were performed using Simplexa ™ Dengue Molecular Assay quantitative real-time RT-PCR [20] performed in a 3M Integrated Cycler machine (Focus Diagnostic, Cypress, CA, USA). Detailed methods for the Simplexa ™ Dengue Molecular Assay were as described by the manufacturer.…”

Retrospective screening of acute undifferentiated fever serum samples with universal flavivirus primers

“…Only 5 of these patients were amongst the PCR positive samples, showing a relatively low sensitivity for virus detection by RT-PCR in serum samples. While optimized RT-PCR methodologies specific for DENV have reported detection rates of over 75% [17], the objective of this study was to survey for different flaviviruses, and so the lower detection rate with a non-optimized protocol for DENV was not surprising. Two of the samples shown to be positive by RT-PCR in this study were not diagnosed as dengue fever patients by serologic and/or NS1 testing [12] suggesting that some cases of dengue fever in Thailand are still missed during clinical screening, possibly as a consequence of .…”