Rapid detection of important human pathogenic Phleboviruses

Weidmann, Manfred; Sánchez-Seco, María Paz; Sall, Amadou Alpha; Ly, Peinda Ogo; Thiongane, Yaya; Lô, Modou Moustapha; Schley, Heike; Hufert, Frank T.

doi:10.1016/j.jcv.2007.10.001

Cited by 99 publications

(76 citation statements)

References 23 publications

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“…For DNA virus, the variola virus (VARV) HA gene was synthesized and ligated into pMA-RQ by Geneart, Regensburg, Germany, and the vaccinia virus (VACV) plasmid carrying the LE gene was provided by the Robert-Koch-Institut (22). For RNA viruses, quantitative Ebola virus (EBOV), Sudan virus (SUDV), and Marburg virus (MARV) NP-gene RNA standards were used as described previously (23,24). A new quantitative sigma virus (SIGV) G gene based RNA standard was generated and transcribed as described previously (24).…”

Section: Methodsmentioning

confidence: 99%

“…For RNA viruses, quantitative Ebola virus (EBOV), Sudan virus (SUDV), and Marburg virus (MARV) NP-gene RNA standards were used as described previously (23,24). A new quantitative sigma virus (SIGV) G gene based RNA standard was generated and transcribed as described previously (24).…”

Section: Methodsmentioning

confidence: 99%

“…(22)(23)(24). A new real-time PCR amplicon was designed for the SIGV G gene and for the VARV HA gene.…”

Section: Methodsmentioning

confidence: 99%

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Development of a Panel of Recombinase Polymerase Amplification Assays for Detection of Biothreat Agents

et al. 2013

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Syndromic panels for infectious disease have been suggested to be of value in point-of-care diagnostics for developing countries and for biodefense. To test the performance of isothermal recombinase polymerase amplification (RPA) assays, we developed a panel of 10 RPAs for biothreat agents. The panel included RPAs for Francisella tularensis, Yersinia pestis, Bacillus anthracis, variola virus, and reverse transcriptase RPA (RT-RPA) assays for Rift Valley fever virus, Ebola virus, Sudan virus, and Marburg virus. Their analytical sensitivities ranged from 16 to 21 molecules detected (probit analysis) for the majority of RPA and RT-RPA assays. A magnetic bead-based total nucleic acid extraction method was combined with the RPAs and tested using inactivated whole organisms spiked into plasma. The RPA showed comparable sensitivities to real-time RCR assays in these extracts. The run times of the assays at 42°C ranged from 6 to 10 min, and they showed no cross-detection of any of the target genomes of the panel nor of the human genome. The RPAs therefore seem suitable for the implementation of syndromic panels onto microfluidic platforms. Syndromic panels for infectious and emerging infectious diseases have been suggested to be of value in point-of-care (POC) diagnostics for developing countries and for biodefense (1). Since the introduction of molecular diagnostics and in particular real-time PCR, ample proof of its sensitivity and specificity has been generated. Indeed, molecular diagnostics are deemed superior to bacterial culture techniques or serological diagnostics (2-4). It has even been suggested to entirely eliminate the old methods in order to streamline centralized laboratories for molecular diagnostics (5-7).In recent years alternative isothermal amplification methods which can be categorized into (i) T7 promoter-driven amplifications (transcription-mediated amplification [TMA], nucleic acid sequence-based amplification [NASBA], and single primer isothermal amplification [SPIA]), (ii) strand displacement methods (strand displacement amplification [SDA], loop-mediated isothermal amplification [LAMP], and smart amplification [SmartAmp]), (iii) helicase-dependent amplification (HDA), (iv) recombinase polymerase amplification (RPA), and (v) rolling-circle amplification (RCA) methods (8-12) have been developed. Some were purposely designed for isothermal amplification starting from RNA (TMA, NASBA, and SPIA), whereas others initially targeted DNA (SDA, LAMP, HDA, RPA, and RCA) and were only later adapted for RNA targets. Nonspecific intercalating fluorophores or fluorescent primers have been used for real-time detection in LAMP, SDA, HDA, and RCA, and specific detection probe formats have been developed for NASBA, RCA, HDA,.In isothermal and exponential RPA, the phage recombinase UvsX and its cofactor UvsY form a nucleoprotein complex with oligonucleotide primers to scan for homologous sequences in a DNA template. Recognition of a specific homologous sequence leads to the initiation of strand invasion of the com...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Development of a Panel of Recombinase Polymerase Amplification Assays for Detection of Biothreat Agents

et al. 2013

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“…There are three genotypes of TOSV, more or less depending upon the geographical origin: lineage A strains were reported in Italy, France, Tunisia and Turkey; lineage B strains originated from Portugal, Spain, France and Morocco [7]; the lineage C was described recently in Croatia and Greece although the virus has not been isolated so far. In all the countries around the Mediterranean Sea TOSV is endemic and should not be considered anymore as an emerging pathogen but rather as a neglected pathogen.…”

Section: Discussionmentioning

confidence: 99%

Afebrile meningoencephalitis with transient central facial paralysis due to Toscana virus infection, south-eastern France, 2014

et al. 2014

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We report a case of meningoencephalitis caused by Toscana virus (TOSV) with central facial paralysis lasting over two days acquired in south-eastern France. The patient was not febrile either before or during the course of the disease. The diagnosis was established by both real-time RT-PCR and virus isolation with complete genome sequencing. This case emphasises the need to consider TOSV in non-febrile neurological syndromes in people living in or having travelled to the Mediterranean area.In this report, we present a case of Toscana virus (TOSV) meningoencephalitis with a central facial paralysis and without fever, acquired in the close vicinity of Marseille, in southern France. This report should be a reminder for healthcare professionals to consider TOSV diagnosis even in afebrile patients with neurological symptoms. Case reportOn 11 July 2014, a French woman in her forties living in a small city at around 20 km from Marseille had a sudden onset of frontal and temporal lobe throbbing headaches upon awakening, without fever. Symptoms were severe and a few hours later she visited the emergency department of a local hospital with headache, nausea and photophonophobia. On admission, she was still non-febrile and the symptoms were persisting despite oral intake of paracetamol in hospital (1,000 mg). The patient's medical history consisted of asthma during childhood, and no history of recent travel abroad.Neurological examination revealed (i) a slight stiff neck, (ii) a discrete left central facial paralysis, and (iii) the absence of abnormalities of cranial nerves except discrete left central facial paresis, sensorimotor loss, cerebellar or pyramidal syndromes. She received 20 mg intramuscular nefopam hydrochloride which alleviated the pain.A lumbar puncture was performed and the analysis showed that the cerebrospinal fluid (CSF) was clear; it contained 475 cells/mm 3 (norm: < 10 cells/mm 3 ) (90% lymphocytes), and showed hypoglycorrhachia at 2.4 mmol/L (norm: > 3mmol/L)) and hyperproteinorachia at 1.78 g/L (norm: 0.4 g/L).A diagnosis of meningoencephalitis was determined. The patient was treated with acyclovir, ceftriaxone and amoxicillin, and was transferred to the neurology department of the same hospital.Further laboratory diagnostic and radiological tests To search for the cause of the meningoencephalitis, a series of laboratory tests were performed. Liver function tests and ionogram were normal. Blood levels of glucose, urea nitrogen, albumin, creatinine, lactates, troponin T, C-reactive protein, and procalcitonin were normal. However, creatinine phosphokinase (CPK) level was moderately increased at 368 U/L (norm: < 170 U/L). There was a slight increase of fibrinogen rate at 4.76 g/L (norm: 2-4 g/L). Prothrombin time was 92% (norm: 70-100%). Blood cell count was 9,100 leukocytes/µL (norm: 4,000-10,000 leukocytes/µL) (81.5% neutrophils and 13.0% lymphocytes), haemoglobin 13.4 g/dL, and the platelet count 251,000/µL.Blood cultures collected on day 1 remained sterile. PCR for Listeria monocytogenes was negative...

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“…For molecular detection of EBOV, the Realstar® Filovirus screen RT-PCR kit (Altona Diagnostics Hamburg, Germany) was available and recommended by WHO (Altona diagnostic, Filovirus real time RT-PCR). The IPD however used quantitative Taqman RT-PCRs for the detection of hemorrhagic fever viruses including Ebola Zaire, Ebola Sudan, Marburg, Yellow fever, Rift valley fever and Crimean Congo on a portable Smartcycler TD machine developed and validated in the context of the EU project VHF Diagnostics (FP6-INCO-032180) [3][4][5][6]. Serological assays were also used for detection of specific IgM and IgG (Winnipeg Canada and CDC USA) for particular cases.…”

Section: Diagnostic Activitiesmentioning

confidence: 99%

The Pasteur Laboratory associated with the Ebola Treatment Center of Macenta (Guinea)

Reynard¹,

Fizet²,

Picard³

et al. 2016

Clin Microbiol Infect Dis

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Rapid detection of important human pathogenic Phleboviruses

Cited by 99 publications

References 23 publications

Development of a Panel of Recombinase Polymerase Amplification Assays for Detection of Biothreat Agents

Development of a Panel of Recombinase Polymerase Amplification Assays for Detection of Biothreat Agents

Afebrile meningoencephalitis with transient central facial paralysis due to Toscana virus infection, south-eastern France, 2014

The Pasteur Laboratory associated with the Ebola Treatment Center of Macenta (Guinea)

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