title: Highly sensitive detection of dengue virus nucleic acid in samples from clinically ill patients.
abstract: Dengue virus (DENV) is a major cause of febrile illness and hemorrhagic fever in tropical and subtropical regions. Typically, patients presenting with acute dengue disease are viremic but may not have yet developed detectable titers of antibody. Therefore, early diagnosis depends mostly on detection of viral components, such as the RNA. To define the potential use of transcription-mediated amplification (TMA) DENV RNA as a diagnostic tool, we first compared its analytic sensitivity using a routine real-time reverse transcription (RT)-PCR and found that TMA is approximately 10 to 100 times more sensitive. In addition, we tested acute-phase serum samples (<5 days post-symptom onset) submitted as part of laboratory-based surveillance in Puerto Rico and determined that among patients with serologically confirmed dengue infection, TMA detected DENV RNA in almost 80% of serum specimens that were negative by the RT-PCR test used for diagnosis and in all specimens with positive RT-PCR results. We conclude that TMA is a highly sensitive method which can detect DENV RNA in approximately 89% of clinical, acute-phase serum specimens.
authors.affiliation: Dengue Branch, Division of Vector-Borne Infectious Disease, Centers for Disease Control and Prevention, San Juan, Puerto Rico.
authors.forename: Jorge L
authors.initials: JL
authors.lastname: Muñoz-Jordán
authors.forename: Cynthia S
authors.initials: CS
authors.lastname: Collins
authors.forename: Edgardo
authors.initials: E
authors.lastname: Vergne
authors.forename: Gilberto A
authors.initials: GA
authors.lastname: Santiago
authors.forename: Lyle
authors.initials: L
authors.lastname: Petersen
authors.forename: Wellington
authors.initials: W
authors.lastname: Sun
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Linnen
mesh_terms: D001769:Blood / Q000821:virology*; D003715:Dengue / Q000175:diagnosis* / Q000821:virology; D003716:Dengue Virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011647:Puerto Rico; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D019595:Severe Dengue / Q000175:diagnosis* / Q000821:virology
chemical_list: D012367:RNA, Viral