CDC DENV-1-4 Real-Time RT-PCR Multiplex Assay
CDC DENV-1-4 rRT-PCR Multiplex Assay for Detection and Serotype Identification of Dengue Virus
Characteristics of the CDC DENV-1-4 rRT-PCR multiplex assay
The CDC DENV-1-4 rRT-PCR multiplex assay is a nucleic acid test (NAT) for detection and typing of dengue virus (DENV) in suspected, symptomatic cases approved by the US Food and Drug Administration (FDA) for use in the United States. This assay detects DENV serotypes 1, 2, 3, or 4 from human serum or plasma collected from human patients with signs and symptoms consistent with dengue infection. The assay is intended to be used as a diagnostic test in patients and is not approved for blood bank screening.
The CDC DENV-1-4 rRT-PCR multiplex assay kit includes:
- Oligonucleotide primers and dual-labeled hydrolysis (Taqman®) probes for in vitro qualitative detection of dengue virus serotypes 1, 2, 3, or 4 from serum or plasma collected from human patients with signs and symptoms consistent with dengue (mild or severe).
- Positive control virus mix, which consists of heat-inactivated DENV-1 Haw, DENV-2 NGC, DENV-3 H87, and DENV-4 H241.
- Human Specimen Control (HSC) is a noninfectious 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 reagent.
The human RNase P RNA (RP) is present in cultured cell material and in most clinical samples and detectable by RT-PCR using the primers and probes provided. The device does not include ancillary reagents.
The CDC DENV-1-4 rRT-PCR multiplex assay can be run in singleplex (each DENV serotype detected in a separate reaction) or in multiplex (the four DENV serotypes are run in the same reaction). These two formats provide equal sensitivity.
Intended use of the CDC DENV-1-4 rRT-PCR multiplex assay
The CDC DENV-1-4 rRT-PCR multiplex 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
- For the identification of DENV serotypes 1, 2, 3, or 4 from viral RNA in serum or plasma (sodium citrate) collected from human patients with dengue
- To provide epidemiologic information for surveillance of circulating dengue viruses
The CDC DENV-1-4 rRT-PCR multiplex assay is not approved for the screening of blood or plasma donors.
- Positive results on the CDC DENV-1-4 rRT-PCR multiplex assay indicate current dengue infection.
- Blood samples from patients suspected of having dengue taken 1-7 days after onset of fever should be tested with this test.
- Negative results obtained with this test do not preclude the diagnosis of dengue and should not be used as the sole basis for treatment or other patient management decisions. If results are negative, IgM testing should be considered.
- If the blood sample is taken from a patient after 7 or more days after the onset of symptoms, laboratory diagnosis is best made using a serologic test for IgM antibody to DENV.
Assay distribution
The CDC DENV-1-4 rRT-PCR multiplex assay is distributed to laboratories with personnel who have training and experience in standardized molecular diagnostic testing procedures and viral diagnosis, and appropriate biosafety equipment and containment. Requests can be sent to DenguePCROrdering@cdc.gov.
Information on the CDC DENV-1-4 rRT-PCR multiplex assay for laboratories in the United States
Materials provided in the CDC DENV-1-4 rRT-PCR multiplex assay
- Box 1: Detection Kit (Primer and Probe Sets)
- Box 2: Positive Control Kit (A mix of heat inactivated DENV-1, -2, -3 and -4 standards)
- Box 3: Human Specimen Extraction Control (HSC)
Click to order the CDC DENV-1-4 rRT-PCR multiplex assay or for technical support.
See package insert for the United States here (print only) [PDF – 53 pages]
Kit: rRT-PCR Enzyme Mastermix Options
Reagent: Invitrogen SuperScript™ III Platinum® reactions One-Step Quantitative RT-PCR System (without Rox)*
- Quantity – 100 reactions
- Catalog No: 11732-020
- Quantity – 500 reactions
- Catalog No: 11732-088
Kit: Nucleic Acid Purification Kit Options
Reagent: Qiagen QIAamp® DSP Viral RNA Mini Kit and Qiagen QIAcube Instrument**
- Quantity – 50 Extractions
- Catalog No: 71904 and 9001292
Reagent: Roche MagNA Pure LC total Nucleic Acid Isolation Kit *
- Quantity – 192 Extractions
- Catalog No: 03 038 505 001
*The CDC DENV-1-4 rRT-PCR multiplex assay test performance requires that only qualified ancillary reagent lots be used with the device in the United States. Any lots not specifically qualified by the CDC-Dengue Branch for use with the CDC DENV-1-4 rRT-PCR multiplex assay are not valid for use with this device and may affect device performance.
** Qiagen QIAamp® DSP Viral RNA Mini Kit (Cat # 71904) is produced under Good Manufacturing Practices (GMP). These RNA extraction kits can be used manually or in combination with the QIAcube Instrument (Cat # 9001292).
Other consumables and minor equipment are required as specified in the package insert of the CDC DENV-1-4 rRT-PCR multiplex assay kit.
Information on the CDC DENV-1-4 rRT-PCR multiplex assay for international laboratories
(countries different from the United States)
Materials provided in the CDC DENV-1-4 rRT-PCR multiplex assay
- Box 1: Detection Kit (Primer and Probe Sets)
- Box 2: Positive Control Kit (A mix of heat inactivated DENV-1, -2, -3 and -4 standards)
- Box 3: Human Specimen Extraction Control (HSC)
See international package insert here (print only) [PDF – 51 pages]
Contact Information, Ordering, and Product Support
For technical and product support and to obtain the CDC DENV-1-4 rRT-PCR multiplex assay and a list of qualified ancillary reagents, please contact the CDC Dengue Branch support team directly.
For ordering and technical support send email to: DenguePCROrdering@cdc.gov.
Include the following information in your message:
- Laboratory name and address
- Qualified contact person
- Phone number
- Email address
- Shipping address