Detection of Yellow Fever 17D Genome in Urine

Domingo, Cristina; Yactayo, Sergio; Agbenu, E; Demanou, Maurice; Schulz, Axel; Daskalow, Katjana; Niedrig, Matthias

doi:10.1128/jcm.01775-10

Cited by 60 publications

(53 citation statements)

References 17 publications

(13 reference statements)

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“…The improved sensitivity and specificity profile led us to apply the RT-qPCR in this work, which targets the highly conserved 5=-NC genomic region of YFV (see Fig. S1 in the supplemental material), to urine samples from 17 healthy YF-17D vaccinees previously studied by a protocol amplifying the NS3 gene (7). The new data corroborated the earlier diagnosis of 9 positive and 8 negative samples.…”

Section: Resultssupporting

confidence: 61%

Advanced Yellow Fever Virus Genome Detection in Point-of-Care Facilities and Reference Laboratories

et al. 2012

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Reported methods for the detection of the yellow fever viral genome are beset by limitations in sensitivity, specificity, strain detection spectra, and suitability to laboratories with simple infrastructure in areas of endemicity. We describe the development of two different approaches affording sensitive and specific detection of the yellow fever genome: a real-time reverse transcriptionquantitative PCR (RT-qPCR) and an isothermal protocol employing the same primer-probe set but based on helicase-dependent amplification technology (RT-tHDA). Both assays were evaluated using yellow fever cell culture supernatants as well as spiked and clinical samples. We demonstrate reliable detection by both assays of different strains of yellow fever virus with improved sensitivity and specificity. The RT-qPCR assay is a powerful tool for reference or diagnostic laboratories with real-time PCR capability, while the isothermal RT-tHDA assay represents a useful alternative to earlier amplification techniques for the molecular diagnosis of yellow fever by field or point-of-care laboratories.

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Section: Resultssupporting

confidence: 61%

Advanced Yellow Fever Virus Genome Detection in Point-of-Care Facilities and Reference Laboratories

et al. 2012

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“…Recent studies report frequent (44%) or prolonged (until 198 d) shedding of vaccine RNA in urine of vaccinated individuals. [35][36] Moreover, the elderly may also have prolonged viremia after vaccination with YFV 17D virus. 37 So, it would be no surprise if some tissues also have RNA positivity for a long time after vaccination.…”

Section: Viremia As a Determinant Of Serious Adverse Reactionsmentioning

confidence: 99%

Serious adverse events associated with yellow fever vaccine

Martins

Leal²,

Homma³

2015

Human Vaccines & Immunotherapeutics

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“…However, this is of little clinical value since infected persons rarely seek medical care during the first phase (3). The demonstration of TBEV RNA in urine samples from two patients during CNS manifestations of varying severity suggests an option for PCR-based diagnosis during at least 1 to 2 weeks after the onset of TBE, as shown in other flavivirus infections (4)(5)(6)(7)(8). These findings argue for prospective studies of TBEV RNA detection in urine in patients with TBE, with the aim of elucidating the value of this body fluid as a sample material for improved diagnosis.…”

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confidence: 99%

Detection of Tick-Borne Encephalitis Virus RNA in Urine

et al. 2014

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b T ick-borne encephalitis (TBE) is a viral zoonosis of increasing incidence in Eurasia, and diagnosis relies mainly on the detection of IgM antibodies in serum. We here present two cases of TBE, of four patients tested, in which we have detected TBE virus (TBEV) RNA in urine with real-time PCR during the encephalitic phase. This observation suggests a new diagnostic opportunity that could be evaluated in a larger cohort of patients.In the first case we report here, 1 week after a tick bite, a 61-yearold woman had high fever, nausea, and fatigue lasting for 5 to 6 days. She was asymptomatic for a week, after which the fever returned along with vomiting and severe headache. After admission, her condition worsened. She was unable to follow instructions, became somnolent and disoriented, and had signs of dysphasia. A cerebrospinal fluid (CSF) sample showed a leukocyte count of 69 ϫ 10 6 /liter (41% polymorphonuclear and 59% mononuclear) and elevated albumin. Specific TBE virus (TBEV) IgM antibodies were detected in serum drawn on day 2 after admission, and the patient seroconverted as regards IgG during the hospital stay. Before nucleic acid extraction using the EasyMag instrument (bioMérieux, Marcy l=Etoile, France), 250-l amounts of serum, CSF, and urine samples were added to 2 ml of lysis buffer and incubated for 10 min at room temperature. TBEV RNA was detected from urine by TaqMan PCR performed as described previously (1) on day 19 after fever onset (Fig. 1). The patient's condition improved, and she was discharged after 10 days of hospitalization.The second case was a 70-year-old woman who reported several tick bites during the preceding weeks and was admitted 2 days after the onset of high fever, nausea, and fatigue. She was on treatment with immunosuppressive drugs (mycophenolate, tacrolimus, and prednisolone) due to kidney transplantation. She developed paresis of the lower extremities and lost the strength in her hands. A CSF sample showed 488 ϫ 10 6 leukocytes/liter (28% polymorphonuclear and 72% mononuclear), elevated albumin and lactate, and a slightly decreased glucose quotient. TBEV IgM antibodies (but not IgG) were detected in serum. On day 8 after admission, the patient became deeply unconscious, had a generalized status epilepticus, and was intubated. Magnetic resonance imaging (MRI) demonstrated high signals in the amygdala and hippocampus. Her neurological condition deteriorated, and she died 11 weeks after admission. TBEV RNA was detected from urine on day 7 and day 16 after onset ( Fig. 1). The RNA quantities allowed sequencing of 10,432 nt from TBEV RNA amplicons, as

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Detection of Yellow Fever 17D Genome in Urine

Cited by 60 publications

References 17 publications

Advanced Yellow Fever Virus Genome Detection in Point-of-Care Facilities and Reference Laboratories

Advanced Yellow Fever Virus Genome Detection in Point-of-Care Facilities and Reference Laboratories

Serious adverse events associated with yellow fever vaccine

Detection of Tick-Borne Encephalitis Virus RNA in Urine

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