María-Paz Sánchez-Seco

Viral infections of the central nervous system in Spain: A prospective study

The aim of the study was to determine the incidence of viruses causing aseptic meningitis, meningoencephalitis, and encephalitis in Spain. This was a prospective study, in collaboration with 17 Spanish hospitals, including 581 cases (CSF from all and sera from 280): meningitis (340), meningoencephalitis (91), encephalitis (76), febrile syndrome (7), other neurological disorders (32), and 35 cases without clinical information. CSF were assayed by PCR for enterovirus (EV), herpesvirus (herpes simplex [HSV], varicella‐zoster [VZV], cytomegalovirus [CMV], Epstein–Barr [EBV], and human herpes virus‐6 [HHV‐6]), mumps (MV), Toscana virus (TOSV), adenovirus (HAdV), lymphocytic choriomeningitis virus (LCMV), West Nile virus (WNV), and rabies. Serology was undertaken when methodology was available. Amongst meningitis cases, 57.1% were characterized; EV was the most frequent (76.8%), followed by VZV (10.3%) and HSV (3.1%; HSV‐1: 1.6%; HSV‐2: 1.0%, HSV non‐typed: 0.5%). Cases due to CMV, EBV, HHV‐6, MV, TOSV, HAdV, and LCMV were also detected. For meningoencephalitis, 40.7% of cases were diagnosed, HSV‐1 (43.2%) and VZV (27.0%) being the most frequent agents, while cases associated with HSV‐2, EV, CMV, MV, and LCMV were also detected. For encephalitis, 27.6% of cases were caused by HSV‐1 (71.4%), VZV (19.1%), or EV (9.5%). Other positive neurological syndromes included cerebellitis (EV and HAdV), seizures (HSV), demyelinating disease (HSV‐1 and HHV‐6), myelopathy (VZV), and polyradiculoneuritis (HSV). No rabies or WNV cases were identified. EVs are the most frequent cause of meningitis, as is HSV for meningoencephalitis and encephalitis. A significant number of cases (42.9% meningitis, 59.3% meningoencephalitis, 72.4% encephalitis) still have no etiological diagnosis. J. Med. Virol. 85:554–562, 2013.

Acute meningitis due to Toscana virus infection among patients from both the Spanish Mediterranean region and the region of Madrid

Toscana virus (TOSV) is a member of the genus Phlebovirus that is transmitted to humans by two different species of sand fly and causes acute aseptic meningitis (AAM) and meningoencephalitis in Central Italy. Fifteen cases of AAM due to TOSV have been found at the Spanish province of Granada, but no data regarding the presence of TOSV-related disease in other regions of Spain have been still reported. A collection of 88 serum and 53 cerebrospinal fluid (CSF) samples taken from 81 selected patients with AAM of unknown aetiology, residing at Madrid or at the southern Mediterranean coast of Spain, was retrospectively studied for presence of TOSV-specific antibodies from both IgG and IgM classes. Anti-TOSV IgG was also investigated in 457 serum samples from healthy individuals, aged 2-60 years, residing at the south of the Region of Madrid. Specific IgM in serum and/or intrathecally produced anti-TOSV IgG were detected in seven patients, three residents from the Mediterranean region and the remainder four from the Region of Madrid. The overall prevalence of anti-TOSV among the healthy population studied was 5%. These results confirm the role of TOSV as an agent causing AAM in the Spanish Mediterranean coast, extend these findings to the central region of the country and suggest that TOSV might be producing infection and neurological disease in every area of Spain harbouring significant populations of the viral vectors.

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.

Detection and monitoring of mosquito flaviviruses in Spain between 2001 and 2005.

Between the years 2001 and 2005, a total of 72,895 female mosquitoes were trapped during their season of abundance, and analyzed. They were sorted into 4,723 pools belonging to 20 Culicidae species from the Anopheles, Aedes, Ochlerotatus, Culex, Culiseta, Coquillettidia, and Uranotaenia genera. The aim was to detect arboviral RNA directly from mosquito homogenates for the genera Alphavirus, Flavivirus, and Phlebovirus. The study formed part of general arbovirus transmission research in four of the most important wetlands in Spain; in the provinces of Girona, Barcelona, Tarragona, and Huelva. The mosquitoes were collected using human bait, CO(2) traps, or light traps, and they were pooled according to date of collection, location, and species. No arboviral RNA from known pathogenic arboviruses was found. However, 111 pools tested positive for unknown mosquito Flavivirus, the only genus detected. The Flavivirus sequences identified were different from all known Flavivirus mosquito viruses, but very close to Kamiti River virus or cell fusing agent virus. The maximum likelihood estimation infection rate (MLE) was calculated for all regions and species. Aedes albopictus had the highest MLE at 47.14, followed by Ae. vexans with 43.67 (over the entire area). These species were followed by Culiseta annulata, with 36.00. The most common species, Ochlerotatus caspius and Culex pipiens, had low MLE values-0.94 and 0.38, respectively-over the area as a whole.

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.

Lymphocytic Choriomeningitis Virus–associated Meningitis, Southern Spain

Lymphocytic choriomeningitis virus (LCMV) was detected in 2 patients with acute meningitis in southern Spain within a 3-year period. Although the prevalence of LCMV infection was low (2 [1.3%] of 159 meningitis patients), it represents 2.9% of all pathogens detected. LCMV is a noteworthy agent of neurologic illness in immunocompetent persons.

Detection and identification of orthopoxviruses using a generic nested PCR followed by sequencing.

Abstract Some orthopoxviruses are considered to be potential biological weapons. After the smallpox eradication campaign ended, routine vaccination was stopped around the world. Consequently, a significant portion of the population is now completely unprotected from infection by variola virus and related orthopoxviruses. Some of the symptoms associated with non-variola infections can be similar to smallpox, causing alert and panic situations. These infections should be considered as real public health concerns, so suitable tools for their differential diagnosis are needed. This study aims to devise a simple and easy-to-perform method that is able to detect and identify any orthopoxvirus that might cause infection in humans. In addition, the similarity of the different genes in the genomes of several species of orthopoxviruses is investigated, and orthopoxvirus-universal primer pairs in the tumour necrosis factor receptor II homologue gene are designed, taking full account of nucleotide similarity. A strategy is devised for their sensitive, rapid and costeffective detection and identification, based on a nested PCR followed by sequencing. The efficacy of the method is tested with samples sent by the European Network of Imported Viral Diseases as part of two external quality control assays. All human orthopoxviruses assayed were detected and identified.

Lymphadenopathy in Patients with Chikungunya Virus Infection Imported from Hispaniola: Case Reports

Chikungunya virus (CHIKV) is currently spreading in the Caribbean and America. Lymphadenopathy, described in infections with other alphaviruses, is not commonly reported in CHIKV infections. Painful lymphadenopathy was found in three of the first six CHIKV infections from the current outbreak diagnosed at a reference center in Madrid, Spain.