Immunocytochemical and Ultrastructural Evidence Supporting That Andes Hantavirus (ANDV) Is Transmitted Person-to-Person Through the Respiratory and/or Salivary Pathways.

In South America Andes hantavirus (ANDV) is hosted by the rodent Oligoryzomys longicaudatus (also known as pygmy rice rat). In humans, ANDV causes Hantavirus Pulmonary Syndrome (HPS), with a fatality rate of about 40%. Epidemiologic and molecular evidence has shown that ANDV can be transmitted from person to person. Sin Nombre hantavirus, occurring in North America, and ANDV are genetically related, and both cause HPS with similar clinical evolution and mortality rate. However, only ANDV is transmitted from person to person. A recent hantavirus outbreak in a small village in Southern Argentine, with 29 HPS cases and 11 deaths has brought to mind that person-to-person transmission continues to be a public health emergency. The present investigation was aimed to understand how does ANDV actually spread between persons. Tissue samples of lung and salivary glands from infected Oligoryzomys longicaudatus and lethal cases of human HPS were investigated by bright field immunocytochemistry, multichannel immunofluorescence, and transmission electron microscopy. The findings are consistent with ANDV infection and replication in the lung alveolar epithelium and macrophages, and in the secretory cells of the submandibular salivary glands. In the lung of infected Oligoryzomys longicaudatus and human cases HPS, the bulk of immunoreactive hantavirus antigens was localized in epithelial cells of the alveolar walls and macrophages. The ultrastructural study supports that in the lung of HPS patients the virus replicates in the alveolar epithelial cells with virus particles being discharged into the alveolar lumen. Virus-like particles were seen within vacuoles of the lung macrophages. Considering that these macrophages can reach the conductive segments of the airways, their expectoration becomes a deadly bullet for ANDV transmission. In the submandibular glands of infected rodents and HPS cases, ANDV antigens were in capillary endothelium, the secretory cells and filling the lumen of the excretory pathway. It is proposed that in patients with HPS caused by ANDV the alveolar epithelium and macrophages would be the gate for the airway spreading of the virus, while the salivary glands are a target for virus replication and an exit pathway through saliva.

Long-Term Study of a Hantavirus Reservoir Population in an Urban Protected Area, Argentina.

Green spaces in urban areas can play a key role in protecting wildlife. However, the presence of wildlife in urban areas can lead to human health risks. Although the presence of the rodent species Oligoryzomys flavescens (hantavirus reservoir) has been recorded in cities of Argentina, its population dynamics in this type of habitat is still unknown. Here, we evaluated: (1) long-term spatial and temporal patterns of O. flavescens abundance and how these patterns were influenced by weather factors and (2) the seroprevalence of hantavirus and the identity of the viral lineage circulating in the population that inhabits the Costanera Sur Ecological Reserve, a protected area in the city of Buenos Aires. Genetic results confirmed that the pathogenic ANDES Central Lechiguanas virus is present in O. flavescens populations inhabiting this urban reserve. Abundance of O. flavescens showed interannual and seasonal fluctuations, with maximum values in winter and spring and minimum ones in summer and autumn. Summers with the highest abundances of O. flavescens were preceded by warmer winters, while winters with lower abundances were preceded by warmer summers. On the other hand, accumulated precipitations in the previous 6 months positively affected winter abundance. These results could help the authorities in charge of the green spaces of Buenos Aires to identify priority areas and times of the year for the implementation of preventive measures that minimize the contact of rodents with visitors. Such measures could be intensified when winters are warmer than normal, and summers are cooler and wetter than normal.

Rodent Abundance and Hantavirus Infection in Protected Area, East-Central Argentina.

We captured 3 hantavirus rodent hosts in Otamendi Natural Reserve, Argentina, during 2007-2012. Hantavirus antibodies were found only in Akodon azarae grass mice, mainly in males and old animals. Higher abundance of this species was associated with warm and rainy weather and high water levels, which peaked after a strong El Niño event.

Viral load of patients with hantavirus pulmonary syndrome in Argentina.

Hantavirus causes severe illness including pneumonia, which leads to hospitalization and often death. At present, there is no specific treatment available. The hantavirus pathogenesis is not well understood, but most likely both virus-mediated and host-mediated mechanisms, are involved. The aim of this study was to correlate viral load in samples of hantavirus pulmonary syndrome cases and hantavirus infected individuals, with clinical epidemiological parameters and disease outcome. The variables that could potentially be related with viral load were analyzed. The retrospective study included 73 cases or household contacts, with different clinical evolution. Viral load was measured by reverse-transcription and real time polymerase chain reaction. There was no statistically significant association between blood viral RNA levels and severity of disease. However, viral load was inversely correlated with IgG response in a statistically significant manner. The level of viral RNA was significantly higher in patients infected with Andes virus South lineage, and was markedly low in persons infected with Laguna Negra virus. These results suggest that the infecting viral genotype is associated with disease severity, and that high viral load is associated with a low specific IgG response. Sex, age and disease severity were not related with viral load. Further investigations increasing strikingly the number of cases and also limiting the variables to be studied are necessary.

Daily Movements and Microhabitat Selection of Hantavirus Reservoirs and Other Sigmodontinae Rodent Species that Inhabit a Protected Natural Area of Argentina.

Abundance, distribution, movement patterns, and habitat selection of a reservoir species influence the dispersal of zoonotic pathogens, and hence, the risk for humans. Movements and microhabitat use of rodent species, and their potential role in the transmission of hantavirus were studied in Otamendi Natural Reserve, Buenos Aires, Argentina. Movement estimators and qualitative characteristics of rodent paths were determined by means of a spool and line device method. Sampling was conducted during November and December 2011, and March, April, June, October, and December 2012. Forty-six Oxymycterus rufus, 41 Akodon azarae, 10 Scapteromys aquaticus and 5 Oligoryzomys flavescens were captured. Movement patterns and distances varied according to sex, habitat type, reproductive season, and body size among species. O. flavescens, reservoir of the etiologic agent of hantavirus pulmonary syndrome in the region, moved short distances, had the most linear paths and did not share paths with other species. A. azarae had an intermediate linearity index, its movements were longer in the highland grassland than in the lowland marsh and the salty grassland, and larger individuals traveled longer distances. O. rufus had the most tortuous paths and the males moved more during the non-breeding season. S. aquaticus movements were associated with habitat type with longer distances traveled in the lowland marsh than in the salty grassland. Hantavirus antibodies were detected in 20% of A. azarae and were not detected in any other species. Seropositive individuals were captured during the breeding season and 85% of them were males. A. azarae moved randomly and shared paths with all the other species, which could promote hantavirus spillover events.

Serological evidence of hantavirus infection in apparently healthy people from rural and slum communities in southern Chile.

Hantavirus disease in America has been recognizable because of its rapid progression in clinical cases, occurrence in previously healthy young adults, and high case fatality rate. Hantavirus disease has been proposed now to define the diversity of clinical manifestations. Since 1995, a total of 902 cases of hantavirus pulmonary syndrome have been reported in Chile, caused by Andes virus (ANDV), with overall fatality of 32%. This report describes the sero-epidemiology of hantavirus in apparently healthy people in rural and urban slum communities from southern Chile. Ten of 934 samples yielded a positive result resulting in a seroprevalence of 1.07% (95% confidence intervals: 0.05%-2.0%). A higher proportion of positive samples was found among individuals from rural villages (1.3%) and slums (1.5%) compared with farms (0.5%). Seropositivity was associated with age (p = 0.011), low education level (p = 0.006) and occupations linked to the household (homemaker, retired, or student) (p = 0.016). No evidence of infection was found in 38 sigmodontinae rodents trapped in the peri-domestic environment. Our findings highlight that exposure risk was associated with less documented risk factors, such as women in slum and rural villages, and the occurrence of infection that may have presented as flu-like illness that did not require medical attention or was misdiagnosed.

Induction of protective immunity in a Syrian hamster model against a cytopathogenic strain of Andes virus.

Andes virus (ANDV) is responsible for the Hantavirus Pulmonary Syndrome cases in Argentina and neighboring countries, with moderate to high case-fatality rates. ANDV has some particular features, which make it unique among other members of the Hantavirus genus such as person-to-person transmission and causing a disease similar to Hantavirus Pulmonary Syndrome in the hamster as an animal model. The kinetics of replication in Vero E6 cells of an ANDV strain isolated in Argentina, called Andes/ARG, was studied. Cytopathic effect and the formation of clear plaques were observed and therefore Andes/ARG could be quantified by classic plaque assay. The Andes/ARG strain was found to be highly lethal in Syrian hamsters allowing experiments to demonstrate the protective potential of vaccines. A recombinant nucleocapsid protein of ANDV induced a long lasting antibody response and protective immunity against a homologous challenge, but to a lower extent against heterologous challenge by the Seoul virus.

Intrathecal antibody production in two cases of yellow fever vaccine associated neurotropic disease in Argentina.

During the period 2007-2008 several epizootics of Yellow fever with dead of monkeys occurred in southeastern Brasil, Paraguay, and northeastern Argentina. In 2008 after a Yellow fever outbreak an exhaustive prevention campaign took place in Argentina using 17D live attenuated Yellow fever vaccine. This vaccine is considered one of the safest live virus vaccines, although serious adverse reactions may occur after vaccination, and vaccine-associated neurotropic disease are reported rarely. The aim of this study was to confirm two serious adverse events associated to Yellow fever vaccine in Argentina, and to describe the analysis performed to assess the origin of specific IgM against Yellow fever virus (YFV) in cerebrospinal fluid (CSF). Both cases coincided with the Yellow fever vaccine-associated neurotropic disease case definition, being clinical diagnosis longitudinal myelitis (case 1) and meningoencephalitis (case 2). Specific YFV antibodies were detected in CSF and serum samples in both cases by IgM antibody-capture ELISA. No other cause of neurological disease was identified. In order to obtain a conclusive diagnosis of central nervous system (CNS) infection the IgM antibody index (AI(IgM) ) was calculated. High AI(IgM) values were found in both cases indicating intrathecal production of antibodies and, therefore, CNS post-vaccinal YFV infection could be definitively associated to YFV vaccination.

Hantavirus pulmonary syndrome: encephalitis caused by virus Andes.

Hemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome (HPS) are rodent-borne emerging diseases caused by members of the genus Hantavirus, family Bunyaviridae. Some species of hantavirus may cause encephalitis, but this is the first report in Andes virus associated to HPS.

Genetic characterization of hantaviruses associated with sigmodontine rodents in an endemic area for hantavirus pulmonary syndrome in southern Brazil.

An ecological assessment of reservoir species was conducted in a rural area (Jaborá) in the mid-west of the state of Santa Catarina in southern Brazil, where hantavirus pulmonary syndrome is endemic, to evaluate the prevalence of hantavirus infection in wild rodents. Blood and tissue samples were collected from 507 rodents during seven field trips from March 2004 to April 2006. Some of the animals were karyotyped to confirm morphological identification. Phylogenetic reconstructions of rodent specimens, based on the mitochondrial DNA cytochrome b gene sequences, were also obtained. Hantavirus antibody was found in 22 (4.3%) of the 507 rodents: 5 Akodon montensis, 2 Akodon paranaensis, 14 Oligoryzomys nigripes, and 1 Sooretamys angouya. Viral RNAs detected in O. nigripes and A. montensis were amplified and sequenced. O. nigripes virus genome was 97.5% (nt) and 98.4% (nt) identical to sequences published for Araucaria (Juquitiba-like) virus based on N and G2 fragment sequences. Viral sequences from A. montensis strain showed 89% and 88% nucleotide identities in a 905-nt fragment of the nucleocapsid (N) protein-coding region of the S segment when it was compared with two other Akodontine rodent-associated viruses from Paraguay, A. montensis and Akodon cursor, respectively. Phylogenetic analysis showed the cocirculation of two genetic hantavirus lineages in the state of Santa Catarina, one from O. nigripes and the other from A. montensis, previously characterized in Brazil and Paraguay, respectively. The hantavirus associated with A. montensis, designed Jaborá virus, represents a distinct phylogenetic lineage among the Brazilian hantaviruses.

Hantavirus pulmonary syndrome in Argentina, 1995-2008.

We report a large case series of hantavirus pulmonary syndrome (HPS) in Argentina that was confirmed by laboratory results from 1995 through 2008. The geographic and temporal distribution of cases by age, sex, fatality rate, and risk factors for HPS was analyzed. A total of 710 cases were unequally distributed among 4 of the 5 Argentine regions. Different case-fatality rates were observed for each affected region, with a maximum rate of 40.5%. The male-to-female ratio for HPS case-patients was 3.7:1.0; the case-fatality rate was significantly higher for women. Agriculture-associated activities were most commonly reported as potential risk factors, especially among men of working age. Although HPS cases occurred predominantly in isolation, we identified 15 clusters in which strong relationships were observed between members, which suggests ongoing but limited person-to-person transmission.

Partial genetic characterization of Seoul hantavirus in rats from Buenos Aires City, Argentina, and generation of a Seoul recombinant nucleoprotein antigen

Hemorrhagic fever with renal syndrome (HFRS) is a severe infectious disease characterized by fever, hemorrhage, renal impairment, and thrombocytopenia. At least seven hantaviruses cause HFRS: Hantaan, Seoul (SEOV) (distributed worldwide), Dobrava/Belgrade, Saaremaa, Amur, Thailand and Puumala. To investigate the epidemiology of HFRS and virus transmission in Argentina, we constructed a prokaryotic plasmid encoding the SEOV rNP, of 430 amino acids. After expression, the rNP was tested as an antigen for use in an enzyme-linked immunosorbent assay for infection diagnosis. To determine the current level of virus transmission in wild brown rats or Norway rats (Rattus norvegicus) captured in Buenos Aires City, Argentina, we tested tissues from rats that were determined to be serologically positive for the SEOV, and the viral genome were detected by RT-PCR using specific primers for two fragments of M segment-encoding Gn and Gc proteins. The viral genome was detected in 11 of 21 seropositive rats (52.4 per cent) captured in two parklands. Sequence analysis of a 333-nt region of the Gc-encoding M segment revealed 97 per cent and 96 per cent identity with strains of SEOV from Baltimore and Brazil, respectively. Our genetic data confirm a very low diversity among SEOV virus strains...

A febre hemorrágica com síndrome renal (FHSR) é uma doença grave, caracterizada por febre, hemorragia, falência renal e trombocitopenia. Pelo menos sete hantavírus causam a FHSR: Hantaan, Seoul (SEOV) (de distribuição global), Dobrava-Belgrade, Saaremaa, Amur, Thailand e Puumala. Para investigar a epidemiologia da FHRS e a transmissão viral na Argentina, criamos um plasmídio procariótico que "codifica" a nucleoproteína recombinante do vírus SEOV de 430 aminoácidos. Após a expressão, a nucleoproteína recombinante foi testada como antígeno para uso em ensaio imunoenzimático (ELISA) para diagnóstico da infecção. Para determinar o nível atual de transmissão viral em populações de ratos-marrons ou ratazanas (Rattus norvegicus) capturadas na cidade de Buenos Aires, Argentina, testamos tecidos de ratos selecionados para serem sorologicamente positivos para o vírus SEOV, e o seu genoma viral foi detectado após submetido a RT-PCR utilizando primers específicos para dois fragmentos de proteínas Gn e Gc codificadas pelo segmento M. O genoma viral foi detectado em 11 das 21 ratazanas soropositivas (52,4 por cento), previamente capturadas em dois parques. A análise sequencial de uma região gênica (333 nt) do segmento M "codificador" da proteína Gc apresentou 97 por cento e 96 por cento de similaridade com as cepas de SEOV coletadas em Baltimore e no Brasil, respectivamente. Os dados genéticos levantados confirmam a informação de que há uma diversidade muito pequena entre as cepas do vírus SEOV...

Expansión geográfica del síndrome pulmonar por hantavirus en la Argentina: Informe del caso más austral / Geographic expansion of hantavirus pulmonary syndrome in Argentina: The southernest case report

Desde 1995 se han informado más de 1000 casos de síndrome pulmonar por hantavirus (SPH) en la Argentina, enfermedad grave y muchas veces fatal para los humanos. La mayoría de los casos fueron asociados al virus Andes (AND) único hantavirus que ha sido informado como causante de transmisión persona a persona. Se han descrito varios linajes patogénicos del virus AND, de los cuales AND Sur, cuyo reservorio es el roedor Oligoryzomys longicaudatus, afecta a la región patagónica de Argentina y Chile. En el presente estudio se informan las manifestaciones clínicas y las características epidemiológicas de un caso de SPH. El objetivo fue describir la presentación clínica del caso, su entorno epidemiológico, el sitio probable de contagio, la variante viral implicada y su relación con los casos más cercanos notificados. Se realizó el seguimiento clínico, el diagnóstico serológico y molecular y la investigación epidemiológica, incluyendo un estudio de la población de roedores reservorios en las áreas involucradas. Se trató de una presentación clásica de SPH moderada, causada por el linaje viral AND sur y su secuencia nucleotídica se comparó con casos del sur argentino y chileno. El caso de hantavirus investigado resultó ser el más austral (48° 46´ 1.2´´ S; 70° 15´ 0´´ O) notificado hasta el momento e involucró a una nueva provincia argentina.

Since 1995 more than 1000 cases of hantavirus pulmonary syndrome (HPS) were reported in Argentina, a severe disease and often fatal to humans. Most cases were associated with Andes virus (AND) that caused few events of person-to-person transmission. Several lineages of pathogenic AND viruses have been described, including AND South, hosted by the rodent Oligoryzomys longicaudatus which affects the Patagonian region of Argentina and Chile. We studied the clinical and epidemiological characteristics of a HPS case. The objective was to describe the clinical presentation of the case, its epidemiology, the likely site of infection, the viral variant implicated and its relationship with the closest reported cases. We carried out the clinical follow up, serological and molecular diagnosis and the epidemiological research, including a rodent reservoir study. The clinical presentation of the case was the classical and moderate, caused by AND South virus. Its viral nucleotide sequence was compared with cases from Southern Argentina and Chile. This case was found to be the most Southern (48° 46´ 1.2´´ S; 70° 15´ O) case reported and involved a new Argentinean province.

Evaluation of an enzyme-linked immunosorbent assay based on Araraquara virus recombinant nucleocapsid protein.

Laboratory diagnosis of hantavirus cardiopulmonary syndrome (HCPS) in Brazil has been performed mostly by detection of IgM antibodies to recombinant antigen purified from Sin Nombre virus and Andes virus (ANDV). Recently, a recombinant nucleocapsid (rN) protein of Araraquara virus (ARAV), a Brazilian hantavirus, was obtained in Escherichia coli. To evaluate ARAV rN as antigen for antibody detection, serum samples from 30 patients from Argentina seropositive for hantavirus were tested. All samples were positive for IgG and IgM by enzyme-linked immunosorbent assay (ELISA) using either ARAV rN or ANDV rN antigens. In Brazil, six of 60 serum samples from patients with suspected HCPS (10%) were positive for IgM by ELISA using ARAV rN antigen and 7 were positive using ANDV rN antigen. For results obtained with 90 serum samples analyzed by IgM ELISA with ANDV rN antigen, the sensitivity of the IgM ELISA using ARAV rN antigen was 97.2%, the specificity was 100%, the positive predictive value was 100%, and the negative predictive value was 98.1%. The results show that ARAV rN is a suitable antigen for diagnosis of hantavirus infection in Brazil and Argentina.

[Geographic expansion of hantavirus pulmonary syndrome in Argentina. The southernest case report]. / Expansión geográfica del síndrome pulmonar por Hantavirus en la Argentina. Informe del caso más austral.

Since 1995 more than 1000 cases of hantavirus pulmonary syndrome (HPS) were reported in Argentina, a severe disease and often fatal to humans. Most cases were associated with Andes virus (AND) that caused few events of person-to-person transmission. Several lineages of pathogenic AND viruses have been described, including AND South, hosted by the rodent Oligoryzomys longicaudatus which affects the Patagonian region of Argentina and Chile. We studied the clinical and epidemiological characteristics of a HPS case. The objective was to describe the clinical presentation of the case, its epidemiology, the likely site of infection, the viral variant implicated and its relationship with the closest reported cases. We carried out the clinical follow up, serological and molecular diagnosis and the epidemiological research, including a rodent reservoir study. The clinical presentation of the case was the classical and moderate, caused by AND South virus. Its viral nucleotide sequence was compared with cases from Southern Argentina and Chile. This case was found to be the most Southern (48 degrees 46' 1.2'' S; 70 degrees 15' O) case reported and involved a new Argentinean province.

Hantavirus antibodies in rodents and human cases with pulmonary syndrome, Rio Negro, Argentina.

In Río Negro Province, Argentina, human cases of hantavirus pulmonary syndrome (HPS) appeared in the region of subantarctic forests. The Andes virus (ANDV) has been identified in the region both in Oligoryzomys longicaudatus rodents and in humans, with the main transmission being from rodents to humans but also showing the possibility of human to human transmission. Between 1996 and 2004, in 40 campaigns, 29.960 night-traps for capturing live rodents were set up. Blood samples were obtained from the rodents and processed using enzyme immunoassay with recombinant antigens made from ANDV. A total of 1767 rodents were captured, with a capture success of 5.9% and an antibody prevalence of 2.1%. Important differences were observed among the species captured from Andes and Steppe regions. Seropositive Oligoryzomys longicaudatus, Abrotrix olivaceus, Abrotrix xanhtothinus and Loxodontomus microtus were captured. During the 1993-2004 period, 40 HPS cases were registered.

Hantavirus antibodies in rodents and human cases with pulmonary syndrome, Río Negro, Argentina / Anticuerpos contra hantavirus en roedores y casos humanos con síndrome pulmonar, Río Negro, Argentina

In Río Negro Province, Argentina, human cases of hantavirus pulmonary syndrome (HPS) appeared in the region of subantartic forests. The Andes virus (ANDV) has been identified in the region both in Oligoryzomys longicaudatus rodents and in humans, with the main transmission being from rodents to humans but also showing the possibility of human to human transmission. Between 1996 and 2004, in 40 campaigns, 29.960 night-traps for capturing live rodents were set up. Blood samples were obtained from the rodents and processed using enzyme immunoassay with recombinant antigens made from ANDV. A total of 1767 rodents were captured, with a capture success of 5.9% and an antibody prevalence of 2.1%. Important differences were observed among the species captured from Andes and Steppe regions. Seropositive Oligoryzomys longicaudatus, Abrotrix olivaceus, Abrotrix xanhtothinus and Loxodontomus microtus were captured. During the 1993-2004 period, 40 HPS cases were registered.

En la Provincia de Río Negro, Argentina, se presentaron casos humanos de síndrome pulmonar por hantavirus (SPH) en la región de los bosques subantárticos. El virus Andes (AND) fue identificado en la región, tanto en el roedor Oligoryzomys longicaudatus como en seres humanos, demostrándose la transmisión principalmente del roedor al hombre y la posibilidad de la transmisión de persona a persona. Para ello, se procedió a la colocación de 29.960 trampas para captura viva de roedores, tipo Sherman, en 40 operativos efectuados desde 1996 hasta 2004. Se obtuvieron muestras de sangre de los roedores, las que fueron procesadas mediante enzimoinmunoensayo con antígenos recombinantes elaborados a partir de virus AND. Fueron capturados 1767 roedores, con un éxito de trampeo del 5.9% y una prevalencia de anticuerpos contra hantavirus del 2.1%. Se observaron importantes diferencias en las especies capturadas en cada una de las regiones. Se capturaron O. longicaudatus, Abrothrix olivaceus y Abrothrix xanhtothinus y Loxodontomys microtus seropositivos. Se registraron 40 casos humanos en el período 1993-2004.

Prevalence of hantavirus infection in wild Rattus norvegicus and R. rattus populations of Buenos Aires City, Argentina.

OBJECTIVE: To determine the prevalence of infection with hantaviruses in wild Rattus norvegicus and R. rattus populations in areas of Buenos Aires city. METHODS: Norway rats and black rats were trapped from 2003 to 2005 at 10 sites throughout Buenos Aires city. Blood samples for serological analysis were collected by cardiac puncture from all individuals captured and tested by enzyme-linked immunosorbent assay for Seoul hantavirus- and Andes virus-specific IgG antibodies. RESULTS: Hantavirus seroprevalence was 11.9% in R. norvegicus (n = 151), varying between 0% and 26.1% depending on the site. Bigger sexually active males were more likely to be infected with Seoul virus than females or juvenile individuals. No antibody-reactive rodents were detected among 33 R. rattus analysed. CONCLUSION: Hantavirus infections are geographically widespread in Buenos Aires city and confirm that they have been enzootic there for at least 20 years.

Pathogenic hantaviruses, northeastern Argentina and eastern Paraguay.

We describe the first, to our knowledge, cases of han-tavirus pulmonary syndrome in northeastern Argentina and eastern Paraguay. Andes and Juquitiba (JUQ) viruses were characterized. JUQV was also confirmed in 5 Oligoryzomys nigripes reservoir species from Misiones. A novel Akodon-borne genetic hantavirus lineage was detected in 1 rodent from the Biologic Reserve of Limoy.

Serological assays for the detection of human andes hantavirus infections based on its yeast-expressed nucleocapsid protein.

BACKGROUND: The objective of the study was to develop and evaluate IgM and IgG ELISAs and an IgG Western blot test for the serological detection of human infections with Andes virus (ANDV), the major cause of hantavirus cardiopulmonary syndrome (HCPS) in South America. METHODS: The entire nucleocapsid (N) protein-encoding sequence of ANDV (strain AH-1) was cloned and expressed in the yeast Saccharomyces cerevisiae. The polyhistidine-tagged recombinant N (rN) protein of ANDV was purified by nickel-chelation chromatography and characterized by its reactivity with different N-specific monoclonal antibodies. To detect an antibody response directed against ANDV in humans, indirect IgM and IgG ELISAs and an IgG Western blot test based on ANDV rN antigen were developed. The evaluation of the tests was performed using a negative serum panel and 63 blinded sera from Argentina and Chile, containing acute-phase and convalescent sera from HCPS patients. RESULTS: The specificities and sensitivities for the IgM and IgG ELISAs were demonstrated to be very high. The IgG ELISA data were confirmed by the IgG Western blot assay based on the same rN antigen. Almost all anti-ANDV-positive sera reacted to higher endpoint titers with N protein of ANDV than with those of Sin Nombre, Laguna Negra or Puumala virus. The cross-reactivity of anti-ANDV-N IgG-positive sera to rN proteins of other hantaviruses was found to be increased with time after the onset of HCPS. CONCLUSION: The high sensitivity of the novel assays should facilitate early diagnosis of ANDV infections and might contribute to a successful treatment of HCPS patients.