Rapid detection of human pathogenic orthobunyaviruses

Modern detection and identification tools can help to provide answers to urgent questions about the incidence, prevalence, and epidemiology of currently emerging diseases. We developed highly sensitive one-step TaqMan reverse transcription-PCR assays with sensitivities ranging from 10(4) to 10(1) mo...

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Vydáno v:Journal of clinical microbiology Ročník 41; číslo 7; s. 3299
Hlavní autoři: Weidmann, Manfred, Rudaz, Veronique, Nunes, Marcio R T, Vasconcelos, Pedro F C, Hufert, Frank T
Médium: Journal Article
Jazyk:angličtina
Vydáno: United States 01.07.2003
Témata:
Animals
Bunyaviridae - classification
Bunyaviridae - genetics
Bunyaviridae - growth & development
Bunyaviridae - isolation & purification
Bunyaviridae Infections - diagnosis
Bunyaviridae Infections - virology
Cell Line
Cercopithecus aethiops
DNA Primers
Humans
Reverse Transcriptase Polymerase Chain Reaction - methods
RNA, Viral - blood
Sensitivity and Specificity
Simbu virus - classification
Simbu virus - genetics
Simbu virus - growth & development
Simbu virus - isolation & purification
Taq Polymerase
Time Factors
Vero Cells
ISSN:0095-1137
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Popis
Shrnutí:Modern detection and identification tools can help to provide answers to urgent questions about the incidence, prevalence, and epidemiology of currently emerging diseases. We developed highly sensitive one-step TaqMan reverse transcription-PCR assays with sensitivities ranging from 10(4) to 10(1) molecules for 11 human pathogens of the orthobunyaviruses. We compared the performances of these assays on three currently available cyclers (ABI-PRISM 7700, LightCycler, and SmartCycler). The assay for Oropouche virus (OROV) was tested using sera collected from days 1 to 5 after onset of OROV disease and was found to be greatly superior to an established nested PCR system. A mean copy number of 1.31 x 10(7) OROV RNA/ml of serum was detected. Diagnostic RNA detection can be used as early as day 1 after onset of OROV disease. The use of a mobile SmartCycler and a hands-on time of less than 3 h could help to intensify outbreak surveillance and control, especially in field studies.
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ISSN:0095-1137
DOI:10.1128/JCM.41.7.3299-3305.2003