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

Santiago, Gilberto A.; Vergne, Edgardo; Quiles, Yashira; Cosme, Joan; Vázquez, Jesús; Medina, Juan F.; Medina, Freddy A.; Colón, Candimar; Margolis, Harold S.; Muñoz‐Jordán, Jorge L.

doi:10.1371/journal.pntd.0002311

Cited by 297 publications

(296 citation statements)

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“…In the United States, Food and Drug Administration (FDA)-approved diagnostic tests for both analytes have been available since 2012. 9,10 A recent highly publicized report by Murray and others 11 describes a retrospective 2009 study looking for evidence of DENV infection in 3,768 serum and CSF specimens from patients in Houston, Texas who were hospitalized during 2003-2005. [11][12][13] This report raises important issues regarding clinical dengue surveillance in the United States and the likelihood of detecting dengue, severe dengue, or dengue with unusual manifestations, such, as neurologic syndromes.…”

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confidence: 99%

Surveillance for Dengue and Dengue-Associated Neurologic Syndromes in the United States

Waterman

Margolis

Sejvar

2015

The American Society of Tropical Medicine and Hygiene

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Abstract. Autochthonous dengue virus transmission has occurred in the continental United States with increased frequency during the last decade; the principal vector, Aedes aegypti, has expanded its geographic distribution in the southern United States. Dengue, a potentially fatal arboviral disease, is underreported, and US clinicians encountering patients with acute febrile illness consistent with dengue are likely to not be fully familiar with dengue diagnosis and management. Recently, investigators suggested that an outbreak of dengue likely occurred in Houston during 2003 based on retrospective laboratory testing of hospitalized cases with encephalitis and aseptic meningitis. Although certain aspects of the Houston testing results and argument for local transmission are doubtful, the report highlights the importance of prospective surveillance for dengue in Aedes-infested areas of the United States, the need for clinical training on dengue and its severe manifestations, and the need for laboratory testing in domestic patients presenting with febrile neurologic illness in these regions to include dengue.Dengue is an arboviral disease of major public health significance caused by four serologically related flaviviruses, dengue viruses (DENVs 1-4), with an expanding worldwide distribution.1 Although many DENV infections are asymptomatic, classic clinical dengue manifests as an acute febrile illness, with symptoms that include headache, retro-orbital pain, myalgia, arthralgia, and rash. Clinical dengue is usually a self-limited illness of about 1 week's duration, but severe illness can occur in both healthy persons and those with other underlying diseases. A relatively small percentage (approximately 10%) of persons with clinical dengue progresses to severe disease because of plasma leakage, which can lead to decompensated shock and death if not carefully managed with appropriate fluid therapy. Most patients requiring hospitalization for dengue have severe disease or complications related to underlying comorbid disease. DENVs seem to be less neurotropic than other flaviviruses, such as West Nile virus, St. Louis encephalitis virus, and Japanese encephalitis virus. DENV-associated neurologic disease, including neuroinvasive disease, has been rarely documented by isolation of virus and detection of DENV-specific immunoglobulin M (IgM) antibody in cerebrospinal fluid (CSF) or brain tissue. In Viet Nam, about 1% of patient hospitalizations with suspected dengue had encephalitis or aseptic meningitis syndromes. 2Another study in Viet Nam found that 0.5% of children admitted for dengue hemorrhagic fever (DHF), which is a subset of severe dengue, had serologically confirmed DENV infection with encephalopathy, although rates of dengue-associated neurologic diseases as high as 21% have been reported. 3,4The most efficient vector for DENV is Aedes aegypti, which is present in limited areas of the southern United States, but its current geographic distribution is not well-documented. Ae. albopictus is a less efficient tran...

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confidence: 99%

Surveillance for Dengue and Dengue-Associated Neurologic Syndromes in the United States

Waterman

Margolis

Sejvar

2015

The American Society of Tropical Medicine and Hygiene

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“…HIV-antibody-negative samples were evaluated for AHI using a p24 antigen assay (miniVidas; bioM erieux) (Sanders et al, 2014). A subset of samples that were negative for both malaria and AHI were tested for DENV infection using RT-PCR (Santiago et al, 2013). All samples from febrile patients that tested positive for malaria or AHI, as well as all samples that tested positive for DENV infection were excluded from the current study except for two samples that were incorrectly included (Fig.…”

Section: Methodsmentioning

confidence: 99%

The plasma virome of febrile adult Kenyans shows frequent parvovirus B19 infections and a novel arbovirus (Kadipiro virus)

Ngoi

Siqueira

et al. 2016

Journal of General Virology

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Viral nucleic acids present in the plasma of 498 Kenyan adults with unexplained fever were characterized by metagenomics analysis of 51 sample pools. The highest to lowest fraction of plasma pools was positive for parvovirus B19 (75 %), pegivirus C (GBV-C) (67 %), alpha anellovirus (59 %), gamma anellovirus (55 %), beta anellovirus (41 %), dengue virus genotype 2 (DENV-2) (16 %), human immunodeficiency virus type 1 (6 %), human herpesvirus 6 (6 %), HBV (4 %), rotavirus (4 %), hepatitis B virus (4 %), rhinovirus C (2 %), Merkel cell polyomavirus (MCPyV; 2 %) and Kadipiro virus (2 %). Ranking by overall percentage of viral reads yielded similar results. Characterization of viral nucleic acids in the plasma of a febrile East African population showed a high frequency of parvovirus B19 and DENV infections and detected a reovirus (Kadipiro virus) previously reported only in Asian Culex mosquitoes, providing a baseline to compare with future virome studies to detect emerging viruses in this region.

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“…The assay was performed using a kit provided by the CDC and following their previously described protocol [15,16]. (Fig.…”

Section: Cdc Denv-1-4 Rrt-pcrmentioning

confidence: 99%

“…These studies have provided details of the genotypes or clades circulating in Thailand [10][11][12]. Although the nested RT-PCR method is more time-consuming and has a higher risk of contamination than realtime RT-PCR (rRT-PCR) [13][14][15][16][17][18][19][20], the cost of the assay is significantly lower making it the preferred method for DENV detection in many laboratories.…”

Section: Introductionmentioning

confidence: 99%

Monitoring and improving the sensitivity of dengue nested RT-PCR used in longitudinal surveillance in Thailand

Klungthong¹,

Manasatienkij²,

Phonpakobsin³

et al. 2015

Journal of Clinical Virology

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a b s t r a c tBackground: AFRIMS longitudinal dengue surveillance in Thailand depends on the nested RT-PCR and the dengue IgM/IgG ELISA. Objective: To examine and improve the sensitivity of the nested RT-PCR using a panel of archived samples collected during dengue surveillance. Study design: A retrospective analysis of 16,454 dengue IgM/IgG ELISA positive cases collected between 2000 and 2013 was done to investigate the sensitivity of the nested RT-PCR. From these cases, 318 acute serum specimens or extracted RNA, previously found to be negative by the nested RT-PCR, were tested using TaqMan real-time RT-PCR (TaqMan rRT-PCR). To improve the sensitivity of nested RT-PCR, we designed a new primer based on nucleotide sequences from contemporary strains found to be positive by the TaqMan rRT-PCR. Sensitivity of the new nested PCR was calculated using a panel of 87 samples collected during 2011-2013. Results and conclusion: The percentage of dengue IgM/IgG ELISA positive cases that were negative by the nested RT-PCR varied from 17% to 42% for all serotypes depending on the year. Using TaqMan rRT-PCR, dengue RNA was detected in 194 (61%) of the 318 acute sera or extracted RNA previously found to be negative by the nested RT-PCR. The newly designed DENV-1 specific primer increased the sensitivity of DENV-1 detection by the nested RT-PCR from 48% to 88%, and of all 4 serotypes from 73% to 87%. These findings demonstrate the impact of genetic diversity and signal erosion on the sensitivity of PCR-based methods.Published by Elsevier B.V.

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

Cited by 297 publications

References 50 publications

Surveillance for Dengue and Dengue-Associated Neurologic Syndromes in the United States

Surveillance for Dengue and Dengue-Associated Neurologic Syndromes in the United States

The plasma virome of febrile adult Kenyans shows frequent parvovirus B19 infections and a novel arbovirus (Kadipiro virus)

Monitoring and improving the sensitivity of dengue nested RT-PCR used in longitudinal surveillance in Thailand

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