CDC DENV-1-4 Real-Time RT-PCR Assay

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Key points

  • Nucleic acid amplification tests (NAAT) can detect dengue virus infections during the first 7 days of illness.
  • CDC's DENV-1-4 Real-Time RT-PCR Assay (DENV-1-4 PCR Assay) is a NAAT for detection and typing of dengue viruses.
  • The DENV-1-4 PCR Assay is approved for the in vitro diagnosis (IVD) of dengue by the U.S. Food and Drug Administration (FDA) and distributed to laboratories.
Three vials of testing solution sitting on ice with a pipette extracting liquid from the one in the center.

Characteristics of the DENV-1-4 PCR Assay

The DENV-1-4 PCR Assay is a NAAT for detection and typing of dengue virus in suspected, symptomatic dengue cases. This assay detects dengue virus serotypes 1, 2, 3, or 4 from serum or plasma collected from human patients.

The DENV-1-4 PCR 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 PR433009, DENV-2 PR347662, DENV-3 PR417637, and DENV-4 PR362977.
  • 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.

The DENV-1-4 PCR Assay can be run in singleplex format (each DENV serotype detected in a separate reaction) or in multiplex format (the four DENV serotypes are mixed in the same reaction). These two formats provide equal sensitivity.

Intended use

The DENV-1-4 PCR Assay is intended for use on an Applied Biosystems (ABI) 7500 Fast Dx Real-Time PCR Instrument for the following purposes:

  • Diagnosis of dengue in serum or plasma collected from patients with signs and symptoms consistent with dengue.
  • 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.

Keep in mind

This assay should be used on specimens collected during the first 7 days after the patient's symptom onset.

The DENV-1-4 PCR Assay is not approved for the screening of blood or plasma donors.

Interpretation of results

  • Positive results on the DENV-1-4 PCR Assay indicate confirmed current dengue infection.
  • Negative results obtained with this test do not rule out the diagnosis of dengue.
  • If the blood sample is taken from a patient after 7 or more days after the onset of symptoms, serologic testing for IgM antibodies to DENV is recommended.
    • IgM antibodies against dengue virus can remain detectable for 3 months or longer after infection.

Assay distribution

The DENV-1-4 PCR 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 for laboratories in the United States

Materials provided in the DENV-1-4 PCR Assay

  • Target-specific primers and probes
  • Positive Controls (A mix of heat inactivated DENV-1, -2, -3 and -4 standards)
  • Human Specimen Extraction Control (HSC)

See package insert for the United States here (print only) [PDF – 53 pages]

Kit: Real-Time RT-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 DENV-1-4 PCR 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 DENV-1-4 PCR 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).

Information on the DENV-1-4 PCR Assay for international laboratories

The DENV-1-4 PCR Assay kits for international laboratories are intended for surveillance of dengue and are labeled as Research Use Only (RUO).

Materials provided in the DENV-1-4 PCR Assay

  • Target-specific primers and probes
  • Positive Controls (A mix of heat inactivated DENV-1, -2, -3 and -4 standards)
  • Human Specimen Extraction Control (HSC)

See international package insert here (print only) [PDF – 51 pages]

Contact information, ordering, and product support

To request the DENV-1-4 PCR Assay kits, a list of qualified ancillary reagents, and to obtain technical assistance, please contact the CDC Dengue Branch support team directly.

For ordering and technical support send an 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
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Content Source:
National Center for Emerging and Zoonotic Infectious Diseases (NCEZID)