Nested RT-PCR for detection of sandfly fever virus, serotype Toscana, in clinical specimens, with confirmation by nucleotide sequence analysis

Schwarz, Tino F.; Jäger, Gundula; Gilch, Sabine; Nitschko, Hans

doi:10.1016/0923-2516(96)80598-x

Cited by 25 publications

(16 citation statements)

References 21 publications

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“…Nevertheless, serological diagnosis is hampered by antigenic cross-reactivity between some members of the group [Tesh et al, 1982;Schwarz et al, 1996b], which may make interpretation of the antibody tests more difficult. In addition, virus isolation is time-consuming, less sensitive, and involves higher biosafety requirements than diagnosis based on viral genome amplification [Schwarz et al, 1995c;Valassina et al, 1996;Braito et al, 1998a]. Molecular methods are often extremely sensitive when reference strains are assayed, but genome variability has to be considered for the design of sensitive diagnostic tests.…”

Section: Introductionmentioning

confidence: 99%

“…More information is, therefore, needed in order to design efficient molecular diagnostic assays. However, some PCR tests have also been developed and have been shown to be useful for diagnosis of TOSV and RVFV infections [Schwarz et al, 1995c;Valassina et al, 1996;Ibrahim et al, 1997;Garcia et al, 2001;Sall et al, 2001]. With the aim of improving the molecular diagnosis of phlebovirus infections, the genetic variability of some Spanish TOSV isolates and of some other phlebovirus strains has been studied.…”

Section: Introductionmentioning

confidence: 99%

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Detection and identification of Toscana and other phleboviruses by RT‐nested‐PCR assays with degenerated primers

Sánchez-Seco¹,

Echevarría²,

Hernández³

et al. 2003

Journal of Medical Virology

152

133

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Phleboviruses are a large and widespread group of viruses that are transmitted by arthropods. Toscana virus is one of the principal agents that causes meningitis in humans during the summer in Italy and, possibly, in other Mediterranean countries. Rift Valley Fever virus can cause serious illness in both animals and humans, leading to high morbidity and mortality, and is considered to be a potential agent for epizootics and human epidemics. Since information on this group of viruses is still scant, reliable laboratory tools for diagnosis and epidemiological surveillance must be developed, in order to ascertain their real impact on Public Health. Sequence data obtained from Spanish isolates of Toscana virus and other phleboviruses confirmed that natural genome variability may hamper the diagnosis of these agents by molecular methods, so this must be borne in mind when developing reliable assays. In view of the above, a novel and useful protocol has been developed for the detection and specific identification of every member of the phlebovirus genus present in a sample, including Toscana virus, based on a generic RT-nested-PCR, followed by sequencing of the amplified fragment. A change in this method also allowed specific direct detection and identification of wild isolates of Toscana virus of different geographical origin, using newly designed primers. Testing clinical samples with these assays confirmed the role of Toscana virus as an agent that causes acute aseptic meningitis in the central region of Spain.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Detection and identification of Toscana and other phleboviruses by RT‐nested‐PCR assays with degenerated primers

Sánchez-Seco¹,

Echevarría²,

Hernández³

et al. 2003

Journal of Medical Virology

152

133

View full text Add to dashboard Cite

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“…Molecular and biological variability among phleboviruses has been studied for RVF virus (Battles & Dalrymple, 1988 ;Anderson & Peters, 1988 ;Saluzzo et al, 1989 ;Besselar et al, 1991) and to a lesser extent for Toscana virus (Schwarz et al, 1995). These reports indicated that among natural isolates of RVF virus, the antigenic properties of the glycoproteins and the nucleoprotein appeared to be stable, and sequences of the G2 epitopes in the M segment are relatively conserved.…”

Section: Introductionmentioning

confidence: 99%

Variability of the NS(S) protein among Rift Valley fever virus isolates.

Sall

Zanotto

Zeller

et al. 1997

Journal of General Virology

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Eighteen strains of Rift Valley fever (RVF) virus collected over a period of 38 years and isolated from diverse localities in Africa and from various hosts (human, animal and arthropod) were investigated by RT-PCR followed by sequencing of the NS S protein coding region. This region was chosen to analyse variability because, in contrast to the N protein, the NS S protein differs in various phleboviruses and there exists an RVF virus (clone 13) in which 70 % of the NS S ORF is deleted, suggesting that this sequence is under a weak selective pressure. Sequence data indicated that percentage di-

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“…Up to 2005, all studies were performed with classic PCR detection based on single round or nested protocols [21,37,56,57] . The description of two genotypes of TOSV supports the need of special attention when designing primers and probes in order to avoid false negative results [58] .…”

mentioning

confidence: 99%

Emergence of Toscana virus in the mediterranean area

Charrel

Bichaud²,

Lamballerie³

2012

WJV

103

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Toscana virus (TOSV) is an arthropod-borne virus, identified in 1971, from Phlebotomus perniciosus and Phlebotomus perfiliewi in central Italy. TOSV belongs to the Phlebovirus genus within the Bunyaviridae family.As other bunyaviruses, the genome of TOSV consists of 3 segments (S for small, M for Medium, and L for Large) respectively encoding non structural and capsid proteins, envelope structural proteins, and the viral RNA-dependant RNA-polymerase. It is transmitted by sand flies. Therefore its distribution is dictated by that of the arthropod vectors, and virus circulation peaks during summertime when sandfly populations are active. Here, we reviewed the epidemiology of TOSV in the old world. First evidence of its pathogenicity for humans, specifically its propensity to cause central nervous system (CNS) infections such as meningitis and encephalitis, was reported in central Italy. After 2000, it was recognized that TOSV had a much larger geographic distribution than initially believed, and was present in most of the Western European countries located on the northern border of the Mediterranean Sea (Portugal, Spain, France, Greece, Croatia) as well as eastern countries such as Cyprus and Turkey. In the countries where TOSV is present, it is among the three most prevalent viruses in meningitis during the warm seasons, together with enteroviruses and herpesviruses. Up to now, epidemiological data concerning Northern Africa and other countries located south of the Mediterranean are scarce. TOSV must be considered an emerging pathogen. Despite the important role played by TOSV in CNS infections, it remains a neglected agent and is rarely considered by physicians in diagnostic algorithms of CNS infections and febrile illness during the warm season, probably because of the lack of information.

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Nested RT-PCR for detection of sandfly fever virus, serotype Toscana, in clinical specimens, with confirmation by nucleotide sequence analysis

Cited by 25 publications

References 21 publications

Detection and identification of Toscana and other phleboviruses by RT‐nested‐PCR assays with degenerated primers

Detection and identification of Toscana and other phleboviruses by RT‐nested‐PCR assays with degenerated primers

Variability of the NS(S) protein among Rift Valley fever virus isolates.

Emergence of Toscana virus in the mediterranean area

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