Correction: Analytical and Clinical Performance of the CDC Real Time RT-PCR Assay for Detection and Typing of Dengue Virus

Abstract

Dengue is an acute illness caused by the positive-strand RNA dengue virus (DENV). There are four genetically distinct DENVs (DENV-1-4) that cause disease in tropical and subtropical countries. Most patients are viremic when they present with symptoms; therefore, RT-PCR has been increasingly used in dengue diagnosis. The CDC DENV-1-4 RT-PCR Assay has been developed as an in-vitro diagnostic platform and was recently approved by the US Food and Drug Administration (FDA) for detection of dengue in patients with signs or symptoms of mild or severe dengue. The primers and probes of this test have been designed to detect currently circulating strains of DENV-1-4 from around the world at comparable sensitivity. In a retrospective study with 102 dengue cases confirmed by IgM anti-DENV seroconversion in the convalescent sample, the RT-PCR Assay detected DENV RNA in 98.04% of the paired acute samples. Using sequencing as a positive indicator, the RT-PCR Assay had a 97.92% positive agreement in 86 suspected dengue patients with a single acute serum sample. After extensive validations, the RT-PCR Assay performance was highly reproducible when evaluated across three independent testing sites, did not produce false positive results for etiologic agents of other febrile illnesses, and was not affected by pathological levels of potentially interfering biomolecules. These results indicate that the CDC DENV-1-4 RT-PCR Assay provides a reliable diagnostic platform capable for confirming dengue in suspected cases.

Natural Infection of Aedes aegypti by Chikungunya and Dengue type 2 Virus in a Transition Area of North-Northeast Brazil

“…The RT-qPCR assay was performed using the SuperScript®III Platinum®One-Step RT-qPCR Kit (ThermoFisher Scientific, Waltham, MA, USA) and the primers and probes used for detection of DENV, ZIKV, and CHIKV [49][50][51]. The reaction was performed on the 7500 Fast real-time PCR thermocycler (ThermoFisher Scientific, Waltham, MA, USA) in a final volume of 25 µL.…”

Development and Application of Performance Assessment Criteria for Next-Generation Sequencing-Based HIV Drug Resistance Assays

“…We identified multiple DRMs at frequencies below 5% that may have been introduced during viral propagation, which suggests infectious clones may not be a suitable reference material for sensitive NGS-based HIVDR assays. Alternatively, in vitro transcription of viral RNA, targeting the Pol region, and spike-ins into an appropriate matrix (plasma, serum, blood) would form high-quality reference materials for quality assessment and has been accomplished with partial or full genomes for other systems [51][52][53][54]. In-vitro-generated viral RNA would provide an excellent source of reference materials that can be used to better define the accuracy and specificity of HIVDR assays.…”

Late-Relapsing Hepatitis after Yellow Fever

“…Markers for autoimmune hepatitis, metabolic liver disease, or infections caused by hepatotropic infectious agents were evaluated (File S1). Urine or sera were used in molecular or serological tests to diagnose infectious diseases [5][6][7][8][9] (File S1). A liver biopsy was collected (93rd DPS) and submitted to histological and immunohistochemistry analyzes, plus qualitative and quantitative molecular investigation of YFV RNA [5,9] (File S1).…”

“…E: Serum and urine were tested by qPCR for YFV[5] and results were negative. Serum was tested by qPCR and negative for DENV[6,7], CHIKV[8], and pan-flaviviruses[9]. Anti-YFV IgM and IgG reagent.…”