Genetic diversity of Toscana virus.

Distribution of Toscana virus (TOSV) is evolving with climate change, and pathogenicity may be higher in nonexposed populations outside areas of current prevalence (Mediterranean Basin). To characterize genetic diversity of TOSV, we determined the coding sequences of isolates from Spain and France. TOSV is more diverse than other well-studied phleboviruses (e.g.,Rift Valley fever virus).

[Clinical laboratory criteria and its utility as predictive of diagnosis of the cardiopulmonary syndrome by hantavirus]. / Criterios de laboratorio clínico y su utilidad como predictores del diagnóstico de síndrome cardiopulmonar por hantavirus.

BACKGROUND: The hantavirus infection is an emerging zoonotic disease, endemic in Chile, generating the hantavirus cardiopulmonary syndrome (HCPS), characterized by cardiopulmonary dysfunction with rapidly progressive respiratory failure and high lethality. For an early clinical orientation of HCPS, due to its non-specificity in symptoms and to help the differential diagnosis, some laboratory parameter that may be useful have been studied. AIM: To identify laboratory criteria as predictive factors of HCPS in patients with suspected hantavirus infection. METHODOLOGY: Retrospective cohort study of 71 patients admitted to the Hospital Guillermo Grant Benavente Emergency. We determined discriminative capacity of laboratory's parameters at the time of admission: platelets recount, hematocrit, inmunoblasts, activated partial thromboplastin time (aPTT) and aspartate aminotransferase (AST/GOT). RESULTS: Were found significant differences in all parameters studied between confirmed patients (22) with respect to unconfirmed (49). Hematocrit, inmunoblasts, AST/GOT and aPTT had a OR > 1 and platelets count had a OR < 1. The best combination for predict HCPS was hematocrit, platelets count and AST/GOT with 90,01% sensibility and 81,63% specificity. CONCLUSION: The five parameters studied are good predictors of HCS in suspicious patients and they would may be useful in low complexity hospitals for quick transfer a center with critical care units.

Reverse transcription, real-time PCR assay for detection of Toscana virus.

BACKGROUND: The arthropod-borne Toscana virus is a common cause of acute neurological infection in the Mediterranean basin. Recently, a new lineage, highly divergent from the Italian prototype, has been reported in Spain. OBJECTIVE: We describe a reverse transcription, real-time PCR assay for detection of both Toscana virus genotypes. The real-time PCR uses a TaqMan probe and an internal control to identify false negative results. STUDY DESIGN: A conserved region of the two known lineages of Toscana virus, located at the 3' end of the small segment of their genomes, was chosen to design both the primers and the probe. RESULTS: The sensitivity of the assay was 0.0158 TICD(50) per reaction of Toscana virus, equivalent to seven copies of cDNA. No other phleboviruses or RNA viruses were amplified by this specific real-time PCR. CONCLUSIONS: The assay seems to be sensitive, reliable and easy to be applied in the diagnosis of autochthonous and/or imported suspected cases of Toscana virus infection.

[Detection of flavivirus in mosquitoes (Diptera: Culicidae) from Easter Island-Chile]. / Detección de flavivirus en mosquitos (Diptera: Culicidae) de la Isla de Pascua-Chile.

Flaviviruses are arthropod-borne viruses, mainly by mosquitoes of the genera Aedes and Culex (Culicidae) that are detected in tropical and subtropical areas. Main flaviviruses of public health importance are: dengue, West Nile virus, yellow fever, among others. In continental Chile, flaviviruses has not been detected. However, there are indigenous cases of dengue detected in Easter Island since 2002, as the presence of its vector Aedes aegypti. The aim of this study was: To determine diversity of flavivirus mosquitoes present in Easter Island. Thirty pools of mosquitoes collected in Hanga Roa were analyzed; a RT-PCR nested flavivirus was performed. Thirteen positive samples were detected and the amplification products were sequenced, identifying two specific flavivirus Insect, the Cell fusing agent virus and other related viruses Kamiti River. This is the first study in Chile showed the presence of flavivirus in vectors in Easter Island.

Criterios de laboratorio clínico y su utilidad como predictores del diagnóstico de síndrome cardiopulmonar por hantavirus / Clinical laboratory criteria and its utility as predictive of diagnosis of the cardiopulmonary syndrome by hantavirus

Resumen Introducción: La infección por hantavirus es una zoonosis emergente, endémica en Chile, generando el síndrome cardiopulmonar por hantavirus (SCPH), caracterizado por disfunción cardiopulmonar con falla respiratoria rápidamente progresiva y altamente letal. Para una orientación clínica precoz del SCPH, debido a su poca especificidad en síntomas y ayudar al diagnóstico diferencial, se han estudiado algunos parámetros de laboratorio que puedan ser de utilidad. Objetivo: Identificar criterios del laboratorio como factores predictores del diagnóstico de SCPH en pacientes con sospecha de enfermedad por hantavirus. Metodología. Estudio de cohorte retrospectiva de 71 pacientes que ingresaron a Urgencia del Hospital Guillermo Grant Benavente. Se determinó la capacidad discriminativa de parámetros de laboratorio al momento de ingreso: recuento de plaquetas, hematocrito, inmunoblastos, TTPa y GOT. Resultados: Se encontraron diferencias significativas en los parámetros estudiados entre pacientes confirmados (n: 22) con respecto a los no confirmados (n: 49). Hematocrito, inmunoblastos, GOT y TTPa tuvieron un OR > 1 y las plaquetas un OR < 1. La mejor combinación para predecir SCPH fue hematocrito, plaquetas y GOT con sensibilidad 90,9% y especificidad 81,6%. Conclusión: Los cinco parámetros estudiados son buenos predictores de SCPH en pacientes con sospecha del mismo y podrían ser útiles en hospitales de baja complejidad para rápido traslado a centro que cuente con unidad de pacientes crítico.

Background. The hantavirus infection is an emerging zoonotic disease, endemic in Chile, generating the hantavirus cardiopulmonary syndrome (HCPS), characterized by cardiopulmonary dysfunction with rapidly progressive respiratory failure and high lethality. For an early clinical orientation of HCPS, due to its non-specificity in symptoms and to help the differential diagnosis, some laboratory parameter that may be useful have been studied. Aim: To identify laboratory criteria as predictive factors of HCPS in patients with suspected hantavirus infection. Methodology: Retrospective cohort study of 71 patients admitted to the Hospital Guillermo Grant Benavente Emergency. We determined discriminative capacity of laboratory's parameters at the time of admission: platelets recount, hematocrit, inmunoblasts, activated partial thromboplastin time (aPTT) and aspartate aminotransferase (AST/GOT). Results: Were found significant differences in all parameters studied between confirmed patients (22) with respect to unconfirmed (49). Hematocrit, inmunoblasts, AST/GOT and aPTT had a OR > 1 and platelets count had a OR < 1. The best combination for predict HCPS was hematocrit, platelets count and AST/GOT with 90,01% sensibility and 81,63% specificity. Conclusion: The five parameters studied are good predictors of HCS in suspicious patients and they would may be useful in low complexity hospitals for quick transfer a center with critical care units.

Different lineages of Chikungunya virus in Equatorial Guinea in 2002 and 2006.

Chikungunya virus is a mosquito-borne virus that causes an acute febrile infection and severe arthralgia and is considered a re-emergent pathogen. During a study investigating arboviruses causing febrile infection in infants in Bata, Equatorial Guinea, the genome of this virus was amplified from blood samples during near two rainy seasons (2002-2003). In 2006, this virus was isolated from a traveler returning to Spain from Equatorial Guinea. These results show that chikungunya virus is present in this country and two lineages are circulating. Thus, this virus should be considered in the differential diagnosis of febrile syndromes in inhabitants and in travelers returning from this country.

Surveillance of arboviruses in Spanish wetlands: detection of new flavi- and phleboviruses.

The presence of viruses in arthropods in Spain has been studied over 5 years. Flaviviruses similar to cell-fusing agent, sequences of a flavivirus related to those transmitted by mosquitoes, and a phlebovirus similar to Naples and Toscana viruses were detected. Their potential human or animal pathogenicity should be studied.

Granada virus: a natural phlebovirus reassortant of the sandfly fever Naples serocomplex with low seroprevalence in humans.

A new member of the phlebovirus genus, tentatively named Granada virus, was detected in sandflies collected in Spain. By showing the presence of specific neutralizing antibodies in human serum collected in Granada, we show that Granada virus infects humans. The analysis of the complete genome of Granada virus revealed that this agent is likely to be a natural reassortant of the recently described Massilia virus (donor of the long and short segments) with a yet unidentified phlebovirus (donor of the medium segment).

Toscana virus in Spain.

Toscana virus (TOSV, Phlebovirus, family Bunyaviridae) infection is one of the most prevalent arboviruses in Spain. Within the objectives of a multidisciplinary network, a study on the epidemiology of TOSV was conducted in Granada, in southern Spain. The overall seroprevalence rate was 24.9%, significantly increasing with age. TOSV was detected in 3 of 103 sandfly pools by viral culture or reverse transcription-polymerase chain reaction from a region of the L gene. Nucleotide sequence homology was 99%-100% in TOSV from vectors and patients and 80%-81% compared to the Italian strain ISS Phl.3. Sequencing of the N gene of TOSV isolates from patients and vectors indicated 87%-88% and 100% homology at the nucleotide and amino acid levels, respectively, compared to the Italian strain. These findings demonstrate the circulation of at least 2 different lineages of TOSV in the Mediterranean basin, the Italian lineage and the Spanish lineage.