Search Results

The CDC DENV-1-4 Real-Time RT-PCR Assay is intended for use on an Applied Biosystems (ABI) 7500 Fast Dx Real-Time PCR Instrument: - For the diagnosis of dengue in serum or plasma collected from patients with signs and symptoms consistent with dengue (mild or severe) during the acute phase: - For the identification of dengue virus serotypes 1, 2, 3 or 4 from viral RNA in serum or plasma (sodium citrate) collected from human patients with dengue during the acute phase; - To provide epidemiologic information for surveillance of circulating dengue viruses.

The CDC DENV-1-4 Real-Time RT-PCR Assay is a panel of oligonucleotide primers and dual-labeled hydrolysis (TaqMan®) probes designed to be utilized in real-time RT-PCR assays using the ABI 7500 Fast Dx Real-Time PCR instrument for detection of DENV-1, -2, -3 and -4 viral RNA in serum or plasma from human patients with dengue. The CDC DENV-1-4 Real-Time RT-PCR Assay includes control materials as described below: - . Positive control materials, which consist of heat-inactivated DENV-1 Haw, DENV-2 NGC, DENV-3 H87, and DENV-4 H241 viruses. - A Human Specimen Control (HSC), consisting of noninfectious (beta ● propiolactone inactivated) cultured human cell material that provides a positive signal in the assay and demonstrates successful recovery of RNA as well as the integrity of the RNA extraction reagents. - . Internal positive control, the human RNase P (RP) primer and probe set detects human RP and is used with each clinical specimen to indicate that adequate isolation of nucleic acid resulted from the extraction of the specimen. A positive RP assay result also ensures that common reagents and equipment are functioning properly and demonstrates the absence of inhibitory substances. RNA extractions can be done using the Qiagen OIAamp® DSP Viral RNA Mini Kit either manually or in combination with the QIAcube Instrument (small centrifuge) and/or Roche MagNA Pure LC total nucleic acid isolation kit and Magna Pure Instrument. The RT-PCR assay is run on the Applied Biosystems® 7500 Fast Dx Real-Time PCR instrument using Invitrogen SuperScript™ III Platinum® One-Step Quantitative RT-PCR System. The assay can be run in singleplex (each DENV serotype detected in a separate reaction) or in multiplex (four DENV serotypes are run in the same reaction). The two formats provide equal sensitivity.