Complete nucleotide sequence of the M RNA segment of Andes virus and analysis of the variability of the termini of the virus S, M and L RNA segments.

Hantavirus pulmonary syndrome (HPS) has been recognized increasingly as a significant public health problem in South America since Andes virus was first discovered in Argentina. Here, the isolation of Andes virus is reported from an infected rodent captured in Argentina in close vicinity to the place of the first HPS case, AH1. The complete nucleotide sequences of the virus M segment, partial L segment and the termini of the S, M and L segment genome RNAs were determined. The Andes virus M RNA segment is 3671 nt in length and is predicted to encode a glycoprotein precursor 1138 aa in length; it generally resembles the other HPS-associated hantaviruses in its organization. Relative to the G1 glycoprotein of other HPS-associated hantaviruses, an additional potential glycosylation site was found but this is located in the predicted cytoplasmic domain and is therefore unlikely to be glycosylated. In phylogenetic analyses, Andes virus, together with the more related hantaviruses, represented a monophyletic lineage. The S-terminal nucleotides were conserved relative to other New World hantaviruses. The M and L segment RNA termini had short deletions in the region believed to contain the sequence and structural features necessary for initiation of virus RNA replication and transcription. Clinical manifestations of Andes virus infections range from fulminant respiratory disease with high lethality to mild course without sequelae. Andes virus has also been associated with person-to-person transmission. Accumulation of Andes virus genetic data will be essential for understanding the factors that regulate virus replication and transmission and to determine the pathogenesis of HPS.

[Hantavirus pulmonary syndrome in southern Argentina]. / Síndrome pulmonar por Hantavirus en el sur andino argentino.

Andes virus was identified in 1995 as the etiologic agent of Hantavirus Pulmonary Syndrome (HPS) in Southern Argentina. We describe herein the main clinical characteristics of 25 HPS confirmed cases acquired in this area between 1993 and September 1999. The mean age was 34 years (range 11-70), with 72% males. Clinical characteristics were similar to those previously reported for Sin Nombre virus (SNV) cases. However, in this group of patients we also observed conjuntival injection in 10/25 (42%), facial flushing in 8/25 (33%), pharyngeal congestion in 7/25 (29%) and petechiae in 3/25 (12%). On the other hand, BUN was increased in 83% of cases (mean 0.77 g/l range 0.31-2.01). Mean serum creatinine concentration was 26.8 mg/l (range: 8.1-110 mg/l) with serum creatinine being higher than 20 mg/l in 8/15 patients (53%). Urinalysis was abnormal in 12/12 cases and was characterized by presence of proteins, red blood cells and granular casts. Aminotransferases were increased in 90% of cases with levels 5-10 times over normal values in 50% of cases. Serum creatine kinase concentration was elevated in 11/14 cases. Two patients required hemodialysis. Case fatality rate was 44% (11/25) and 10 of these cases died among the first 10 days of illness. Mononuclear myocarditis was observed in two cases, a finding that has not been reported for SNV cases. During the 1996 HPS outbreak in Southern Argentina due to Andes virus, there were epidemiological and molecular evidences of person-to-person transmission, a feature not previously shown for other members of the hantavirus genus. These data would also be indicative of some distinctive clinical characteristics of HPS caused by Andes virus, with more frequent renal involvement than in SNV cases.

Genetic diversity, distribution, and serological features of hantavirus infection in five countries in South America.

Since 1995 when the first case of hantavirus pulmonary syndrome (HPS) was reported in Patagonia, there have been more than 400 cases of HPS reported in five countries in South America. The first case of HPS was associated with Andes (AND) virus. In this study, we report on the genetic diversity, geographical distribution, and serological features of hantavirus infection in six countries in South America based on 87 HPS cases from Argentina, Bolivia, Chile, Paraguay, and Uruguay. An early immunoglobulin M (IgM), IgA, and IgG humoral response was observed in almost all HPS cases. The IgM response appears to peak 1 or 2 days after the onset of symptoms. Peak IgG antibody titers occur mostly after the first week. Low IgG titers or the absence of IgG was associated with higher mortality rates. The IgA response peaks around day 15 and then rapidly decreases. The results of phylogenetic analysis based on partial M-fragment G1- and G2-encoding sequences showed that HPS cases from the five countries were infected with viruses related to AND or Laguna Negra (LN) virus. Within AND virus-infected persons, at least five major genetic lineages were found; one lineage was detected in Uruguayan and Argentinean cases from both sides of the Rio de la Plata river. Two Paraguayan patients were infected with a virus different from LN virus. According to the results of phylogenetic analyses, this virus probably belongs to a distinct lineage related more closely to the AND virus than to the LN virus, suggesting that there is probably an Oligoryzomys-borne viral variant circulating in Paraguay. These studies may contribute to a better understanding of hantavirus human infection in South America.

Development and evaluation of a solid-phase enzyme immunoassay based on Andes hantavirus recombinant nucleoprotein.

Hantavirus pulmonary syndrome (HPS) with high mortality rate has been reported in five countries in South America. Rapid accurate methods are important both for monitoring acute infections and for epidemiological studies. The Andes virus nucleoprotein amino acid sequence has a high identity percentage compared with other sequences of this region and has been chosen for the development of diagnostic reagents. Andes nucleoprotein expressed in Escherichia coli was applied as antigen in IgG, IgA and mu-capture IgM enzyme-linked inmunosorbent assays (ELISAs). An evaluation of this reagent was conducted to establish its usefulness for differential diagnosis of HPS and seroprevalence studies. Samples from 135 reverse transcription (RT)-PCR-confirmed HPS cases, 77 individuals with other respiratory infections and 957 healthy inhabitants from endemic and non-endemic areas were analysed. The hantavirus-infected patients had an early and strong IgM, IgG and IgA serum antibody response, in most of the cases as early as 1, 7 and 1 days following onset of symptoms, respectively. IgM and IgG detection showed a specificity and sensitivity of 100%. Andes-specific IgM antibodies were found in all patients in the first available sample, which remained detectable for at least 43 days. Specific IgA antibodies were also detected in saliva of patients with acute HPS. The short duration of the disease and the risk for contacts due to person-to-person transmission of Andes virus necessitate the use of highly sensitive tests which might lead to earlier detection of infected people and improve the treatment and management of patients with HPS.

Síndrome pulmonar por Hantavirus en el sur andino argentino. / [Hantavirus pulmonary syndrome in southern Argentina]

Andes virus was identified in 1995 as the etiologic agent of Hantavirus Pulmonary Syndrome (HPS) in Southern Argentina. We describe herein the main clinical characteristics of 25 HPS confirmed cases acquired in this area between 1993 and September 1999. The mean age was 34 years (range 11-70), with 72

males. Clinical characteristics were similar to those previously reported for Sin Nombre virus (SNV) cases. However, in this group of patients we also observed conjuntival injection in 10/25 (42

), facial flushing in 8/25 (33

), pharyngeal congestion in 7/25 (29

) and petechiae in 3/25 (12

). On the other hand, BUN was increased in 83

of cases (mean 0.77 g/l range 0.31-2.01). Mean serum creatinine concentration was 26.8 mg/l (range: 8.1-110 mg/l) with serum creatinine being higher than 20 mg/l in 8/15 patients (53

). Urinalysis was abnormal in 12/12 cases and was characterized by presence of proteins, red blood cells and granular casts. Aminotransferases were increased in 90

of cases with levels 5-10 times over normal values in 50

of cases. Serum creatine kinase concentration was elevated in 11/14 cases. Two patients required hemodialysis. Case fatality rate was 44

(11/25) and 10 of these cases died among the first 10 days of illness. Mononuclear myocarditis was observed in two cases, a finding that has not been reported for SNV cases. During the 1996 HPS outbreak in Southern Argentina due to Andes virus, there were epidemiological and molecular evidences of person-to-person transmission, a feature not previously shown for other members of the hantavirus genus. These data would also be indicative of some distinctive clinical characteristics of HPS caused by Andes virus, with more frequent renal involvement than in SNV cases.

Hantavirus pulmonary syndrome outbreak in Argentina: molecular evidence for person-to-person transmission of Andes virus.

An increase of Hantavirus Pulmonary Syndrome (HPS) cases around a southwestern Argentina town and in persons living 1400 km away but in contact with those cases was detected during the spring of 1996. In order to evaluate person-to-person transmission we compared the homology of PCR-amplified viral sequences of 26 Argentine and Chilean cases. Sixteen of them were epidemiologically linked cases and had the same sequence (Epilink/96) in the S segment 3' noncoding region and in the M segment partial G1 and G2 region (a total of 1075 nucleotides). Contrarily, two geographical and contemporary but nonepidemiologically related cases differed from Epilink/96 in the compared regions. No significant differences, such as glycosylation or hydrophilic pattern, were found between Epilink/96 and the other sequences. Nucleotide and deduced amino acid sequence homologies between samples from southern Argentina and Chile ranged from 90.9 to 100% and 96.4 to 100%, respectively. Phylogenetic analysis revealed that all the analyzed southwestern viruses belong to the Andes lineage. Although human infection principally occurs via inhalation of contaminated rodent excreta, our results with Andes virus show the first direct genetic evidence of person-to-person transmission of a hantavirus.

Replication of Junin virus in the presence of tunicamycin.

In the presence of tunicamycin (TM), an antibiotic which inhibits glycosylation, Junin virus-infected cells released 70% as much virus as control cultures, as measured by [35S]-methionine appearing in a sucrose gradient virus particle fraction. In this same fraction, the incorporation of [14C]-glucosamine was inhibited 85% by TM, accompanied by a dramatic decrease in the envelope glycoprotein, Gp38, and a marked decrease in virus infectivity. These results indicate that the presence of Gp38 on the viral surface is essential for viral infectivity. In contrast, maturation and release of virus particles do not seem to be affected by the absence of Gp38 from the viral envelope.