Comparison of four reverse transcription-polymerase chain reaction procedures for the detection of dengue virus in clinical specimens

Raengsakulrach, Boonyos; Nisalak, Ananda; Maneekarn, Niwat; Yenchitsomanus, Pa‐thai; Limsomwong, Chandhana; Jairungsri, Aroonroong; Thirawuth, Vipa; Green, Sharone; Kalayanarooj, Siripen; Suntayakorn, Saroj; Sittisombut, Nopporn; Malasit, Prida; Vaughn, David W.

doi:10.1016/s0166-0934(02)00104-0

Cited by 40 publications

(35 citation statements)

References 39 publications

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“…This was despite the fact that acute serum collection occurred at a median of 4.5 days after illness onset, a period when PCR is considered more sensitive than serology. 39,40 Possible explanations for this observation include cold chain breaks during sample transport that would likely have affected PCR results more than serological results or serological cross-reactivity resulting in false anti-DENV IgM positives. Although a positive anti-DENV IgM result in the setting of an acute febrile illness is often considered diagnostic of acute DENV infection, this approach can be problematic because anti-DENV IgM is often detectable for 2-3 months after initial infection.…”

Section: Discussionmentioning

confidence: 99%

Chikungunya and Dengue Fever among Hospitalized Febrile Patients in Northern Tanzania

Hertz

Munishi²,

Ooi³

et al. 2012

The American Society of Tropical Medicine and Hygiene

122

129

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Abstract. Consecutive febrile admissions were enrolled at two hospitals in Moshi, Tanzania. Confirmed acute Chikungunya virus (CHIKV), Dengue virus (DENV), and flavivirus infection were defined as a positive polymerase chain reaction (PCR) result. Presumptive acute DENV infection was defined as a positive anti-DENV immunoglobulin M (IgM) enzyme-linked immunsorbent assay (ELISA) result, and prior flavivirus exposure was defined as a positive anti-DENV IgG ELISA result. Among 870 participants, PCR testing was performed on 700 (80.5%). Of these, 55 (7.9%) had confirmed acute CHIKV infection, whereas no participants had confirmed acute DENV or flavivirus infection. Anti-DENV IgM serologic testing was performed for 747 (85.9%) participants, and of these 71 (9.5%) had presumptive acute DENV infection. Anti-DENV IgG serologic testing was performed for 751 (86.3%) participants, and of these 80 (10.7%) had prior flavivirus exposure. CHIKV infection was more common among infants and children than adults and adolescents (odds ratio [OR] 1.9, P = 0.026) and among HIV-infected patients with severe immunosuppression (OR 10.5, P = 0.007). CHIKV infection is an important but unrecognized cause of febrile illness in northern Tanzania. DENV or other closely related flaviviruses are likely also circulating.

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Section: Discussionmentioning

confidence: 99%

Chikungunya and Dengue Fever among Hospitalized Febrile Patients in Northern Tanzania

Hertz

Munishi²,

Ooi³

et al. 2012

The American Society of Tropical Medicine and Hygiene

122

129

View full text Add to dashboard Cite

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“…The products of these reactions are separated by electrophoresis on an agarose gel, which allows the dengue serotypes to be differentiated on the basis of size. The sensitivity of RT-PCR assays in comparison to virus isolation in mosquito cell culture varies between 25% and 79% 98 .…”

Section: Reverse Transcriptase Pcr (Rt-pcr)mentioning

confidence: 99%

Dengue: a continuing global threat

et al. 2010

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Dengue fever and dengue haemorrhagic fever are important arthropod-borne viral diseases. Each year, there are ~50 million dengue infections and ~500,000 individuals are hospitalized with dengue haemorrhagic fever, mainly in Southeast Asia, the Pacific and the Americas. Illness is produced by any of the four dengue virus serotypes. A global strategy aimed at increasing the capacity for surveillance and outbreak response, changing behaviours and reducing the disease burden using integrated vector management in conjunction with early and accurate diagnosis has been advocated. Antiviral drugs and vaccines that are currently under development could also make an important contribution to dengue control in the future.Dengue is the most important arthropod-borne viral infection of humans. Worldwide, an estimated 2.5 billion people are at risk of infection, approximately 975 million of whom live in urban areas in tropical and sub-tropical countries in Southeast Asia, the Pacific and the Americas 1 . Transmission also occurs in Africa and the Eastern Mediterranean, and rural Europe PMC Funders GroupAuthor Manuscript Nat Rev Microbiol. Author manuscript; available in PMC 2015 February 19. Published in final edited form as:Nat Rev Microbiol. 2010 December ; 8(12 0): S7-16. doi:10.1038/nrmicro2460. Europe PMC Funders Author ManuscriptsEurope PMC Funders Author Manuscripts communities are increasingly being affected. It is estimated that more than 50 million infections occur each year, including 500,000 hospitalizations for dengue haemorrhagic fever, mainly among children, with the case fatality rate exceeding 5% in some areas [1][2][3][4] . The geographical areas in which dengue transmission occurs have expanded in recent years (FIG. 1), and all four dengue virus serotypes (DENV-1-4) are now circulating in Asia, Africa and the Americas, a dramatically different scenario from that which prevailed 20 or 30 years ago (FIG. 2). The molecular epidemiology of these serotypes has been studied in an attempt to understand their evolutionary relationships 11 .This Review will provide an update on our understanding of the pathogenesis of this successful pathogen, how we diagnose and control infection and the progress that has been made in vaccine development. Dengue virus pathogenesisDengue viruses belong to the genus flavivirus within the Flaviviridae family. DENV-1-4 evolved in non-human primates from a common ancestor and each entered the urban cycle independently an estimated 500-1,000 years ago 12 . The virion comprises a spherical particle, 40-50 nm in diameter, with a lipopolysaccharide envelope. The positive singlestrand RNA genome (FIG. 3), which is approximately 11 kb in length, has a single open reading frame that encodes three structural proteins -the capsid (C), membrane (M) and envelope (E) glycoproteins -and seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5 [18][19][20] . ADE occurs when mononuclear phagocytes are infected through their Fc receptors by immune complexes that form between DE...

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“…Reverse transcription (RT)-PCR approaches are being increasingly used for dengue diagnosis with acute-phase serum samples (9,15). The sensitivity of RT-PCR assays in serologically confirmed cases ranges from 40 to 80% and may decrease as the interval from symptom onset to specimen collection increases (23,29). The sensitivity of molecular diagnostics may be affected by such factors as the temperature of the sample during transport and storage, RNA extraction procedures, or the method of detection of PCR products.…”

mentioning

confidence: 99%

Highly Sensitive Detection of Dengue Virus Nucleic Acid in Samples from Clinically Ill Patients

Muñoz‐Jordán

Collins²,

Vergne

et al. 2009

J Clin Microbiol

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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.

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Comparison of four reverse transcription-polymerase chain reaction procedures for the detection of dengue virus in clinical specimens

Cited by 40 publications

References 39 publications

Chikungunya and Dengue Fever among Hospitalized Febrile Patients in Northern Tanzania

Chikungunya and Dengue Fever among Hospitalized Febrile Patients in Northern Tanzania

Dengue: a continuing global threat

Highly Sensitive Detection of Dengue Virus Nucleic Acid in Samples from Clinically Ill Patients

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