ABSTRACT
We conducted a development and standardization of an IgG ELISA assay for serological detection of human orthohantavirus infections using the recombinant antigen rLECH13 produced in bacterial and derived from the LECHV. The evaluation and standardization were carried out by analyzing serum samples from a total of 50 patients with confirmed Hantavirus Pulmonary Syndrome (HPS) diagnosis through the reference technique, 50 negative sera, and 53 patients with other medical conditions. The data from the assay analysis showed a diagnostic sensitivity value of 95% and a diagnostic specificity of 80%. The high sensitivity of this novel assay leads us to conclude that rLECH13 is a feasible option for use in the immunodiagnostic of orthohantavirus infection. Additionally, it is crucial to have an antigen that can be produced under conditions that do not require highly complex laboratories. Furthermore, the new assay is cost-effective, reproducible, and demonstrates excellent performance.
Subject(s)
Antibodies, Viral , Enzyme-Linked Immunosorbent Assay , Hantavirus Infections , Orthohantavirus , Sensitivity and Specificity , Humans , Enzyme-Linked Immunosorbent Assay/methods , Orthohantavirus/immunology , Orthohantavirus/genetics , Orthohantavirus/isolation & purification , Argentina , Hantavirus Infections/diagnosis , Antibodies, Viral/blood , Nucleocapsid Proteins/immunology , Nucleocapsid Proteins/genetics , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Immunoglobulin G/blood , Antigens, ViralABSTRACT
Following an Argentine Hemorrhagic Fever (AHF) outbreak in the early 1990s, a rodent survey for Junín virus, a New World Clade B arenavirus, in endemic areas of Argentina was conducted. Since 1990, INEVH has been developing eco-epidemiological surveillance of rodents, inside and outside the Argentine Hemorrhagic Fever endemic area. Samples from rodents captured between 1993 and 2019 that were positive for Arenavirus infection underwent Sanger and unbiased, Illumina-based high-throughput sequencing, which yielded 5 complete and 88 partial Mammarenaviruses genomes. Previously, 11 genomes representing four species of New World arenavirus Clade C existed in public records. This work has generated 13 novel genomes, expanding the New World arenavirus Clade C to 24 total genomes. Additionally, two genomes exhibit sufficient genetic diversity to be considered a new species, as per ICTV guidelines (proposed name Mammarenavirus vellosense). The 13 novel genomes exhibited reassortment between the small and large segments in New World Mammarenaviruses. This work demonstrates that Clade C Mammarenavirus infections circulate broadly among Necromys species in the Argentine Hemorrhagic Fever endemic area; however, the risk for Clade C Mammarenavirus human infection is currently unknown.
Subject(s)
Arenaviridae , Arenavirus , Arenaviruses, New World , Hemorrhagic Fever, American , Junin virus , Animals , Humans , Arenaviridae/genetics , Rodentia , Hemorrhagic Fever, American/epidemiology , Argentina/epidemiology , Arenaviruses, New World/genetics , Junin virus/genetics , Arenavirus/geneticsABSTRACT
Resumen Desde la identificación del virus Junin en la década del 50, se realizaron numerosos estudios en roedores silvestres dentro del área endémica de la Fiebre Hemorrágica Argentina (FHA) que per mitieron registrar, además, actividad del virus de la coriomeningitis linfocitaria (LCMV) y del virus Latino (LATV). La ausencia de casos confirmados de FHA desde la década del 90 en el departamento Río Cuarto, provincia de Córdoba, promovió la vigilancia ecoepidemiológica y de infección del Calomys musculinus (reservorio del virus Junin) y la búsqueda de reservorios e infección de los otros mammarenavirus. Durante dos años de muestreo estacional, con un sistema de captura, marcación y liberación capturamos 857 roedores, que correspondieron 57.3% a los reservorios: C. musculinus (especie más abundante), C. venustus y Mus musculus. Detectamos anticuerpos y caracterizamos molecularmente los tres agentes virales. Observamos una prevalencia de infección de 3.5% (9/254) para virus Junin, 100% (3/3) para LCMV y 24.1% (21/87) para LATV. En conclusión, demostra mos circulación de virus Junin en su roedor reservorio, en una región considerada histórica para FHA con riesgo potencial para la población y cocirculación espacio-temporal de los tres mammarenavirus en la región central de Argentina.
Abstract Since the identification of Junin virus in the 1950s, many studies were carried out in wild rodents within the endemic area of the Argentine Hemorrhagic Fe ver (AHF) that recorded also the activity of the lymphocytic choriomeningitis virus (LCMV) and the Latino virus (LATV). The absence of confirmed cases of AHF since the 1990s in the department of Rio Cuarto, Córdoba province, promoted ecoepidemiological surveillance of infection of Calomys musculinus (Junin virus reservoir) and the search of reservoirs of the other mammarenaviruses. During two years of seasonal sampling, with a capture, mark and release system, 857 rodents were captured, corresponding 57.3% to the rodent reservoirs: C. musculinus, C. venustus and Mus musculus, being the first the most abundant species. Antibodies were detected and the three viral agents were molecularly characterized, showing a prevalence of infection of 3.5% (9/254) for Junin virus, 100% (3/3) for LCMV and 24.1% (21/87) for LATV. In conclusion, we demonstrated Junin virus circulation in its rodent reservoir in a region considered historic for AHF with potential risk for the population and the spatio-temporal co-circulation of the three mammarenaviruses in the central region of Argentina.
ABSTRACT
Since the identification of Junin virus in the 1950s, many studies were carried out in wild rodents within the endemic area of the Argentine Hemorrhagic Fever (AHF) that recorded also the activity of the lymphocytic choriomeningitis virus (LCMV) and the Latino virus (LATV). The absence of confirmed cases of AHF since the 1990s in the department of Rio Cuarto, Córdoba province, promoted ecoepidemiological surveillance of infection of Calomys musculinus (Junin virus reservoir) and the search of reservoirs of the other mammarenaviruses. During two years of seasonal sampling, with a capture, mark and release system, 857 rodents were captured, corresponding 57.3% to the rodent reservoirs: C. musculinus, C. venustus and Mus musculus, being the first the most abundant species. Antibodies were detected and the three viral agents were molecularly characterized, showing a prevalence of infection of 3.5% (9/254) for Junin virus, 100% (3/3) for LCMV and 24.1% (21/87) for LATV. In conclusion, we demonstrated Junin virus circulation in its rodent reservoir in a region considered historic for AHF with potential risk for the population and the spatio-temporal co-circulation of the three mammarenaviruses in the central region of Argentina.
Desde la identificación del virus Junin en la década del 50, se realizaron numerosos estudios en roedores silvestres dentro del área endémica de la Fiebre Hemorrágica Argentina (FHA) que permitieron registrar, además, actividad del virus de la coriomeningitis linfocitaria (LCMV) y del virus Latino (LATV). La ausencia de casos confirmados de FHA desde la década del 90 en el departamento Río Cuarto, provincia de Córdoba, promovió la vigilancia ecoepidemiológica y de infección del Calomys musculinus (reservorio del virus Junin) y la búsqueda de reservorios e infección de los otros mammarenavirus. Durante dos años de muestreo estacional, con un sistema de captura, marcación y liberación capturamos 857 roedores, que correspondieron 57.3% a los reservorios: C. musculinus (especie más abundante), C. venustus y Mus musculus. Detectamos anticuerpos y caracterizamos molecularmente los tres agentes virales. Observamos una prevalencia de infección de 3.5% (9/254) para virus Junin, 100% (3/3) para LCMV y 24.1% (21/87) para LATV. En conclusión, demostramos circulación de virus Junin en su roedor reservorio, en una región considerada histórica para FHA con riesgo potencial para la población y cocirculación espacio-temporal de los tres mammarenavirus en la región central de Argentina.
Subject(s)
Arenaviridae , Arenaviruses, New World , Hemorrhagic Fever, American , Junin virus , Animals , Disease Reservoirs , Hemorrhagic Fever, American/epidemiology , Humans , Mice , RodentiaABSTRACT
Arenavirus Sabiá was originally isolated from a fatal human infection in Brazil, and after the occurrence of the second fatal human case in São Paulo state, epidemiologic and virologic studies were performed in the area where the patient lived, aiming at the identification of the Sabiá natural rodent reservoir. A broadly cross-reactive enzyme-linked immunosorbent assay (ELISA) was used to screen for antibody-positive samples. Antibodies to arenavirus were detected in two of the 55 samples of Calomys tener, and from these results, samples of rodents were analyzed by a broad RT-PCR assay. RT-PCR amplification detected arenavirus sequences in five of the 55 C. tener samples, and sequencing showed that this virus is a distinct form of Sabiá virus. Thus, we describe here the evidence for the circulation of a new arenavirus in Brazil (proposed name Pinhal virus) and its genetic characterization compared to other arenaviruses. This study also suggests C. tener as a probable rodent reservoir for this virus and associates this new virus with the lineage C of New World arenaviruses. Although we have defined some characteristics of this virus, so far, there is no evidence of its involvement in human disease.
Subject(s)
Arenaviridae Infections/veterinary , Arenaviruses, New World/isolation & purification , Sigmodontinae/virology , Animals , Antibodies, Viral/blood , Arenaviridae Infections/virology , Arenaviruses, New World/classification , Arenaviruses, New World/genetics , Arenaviruses, New World/immunology , Brazil/epidemiology , Disease Reservoirs/veterinary , Enzyme-Linked Immunosorbent Assay , PhylogenyABSTRACT
The complete nucleotide sequences of two West Nile virus (WNV) strains isolated in Argentina were determined. Phylogenetic trees were constructed from the aligned nucleic acid sequences of these two strains along with other previously published complete WNV genome sequences. Phylogenetic data showed that both strains belonged to clade 1a of lineage 1 and clustered in a subclade with American strains isolated during 1999-2002. These results suggest two independent routes of introduction of WNV in Argentina and that the virus could have been circulating in Argentina for some time before being isolated.
Subject(s)
Genome, Viral/genetics , West Nile Fever/veterinary , West Nile virus/genetics , Amino Acid Sequence , Amino Acid Substitution , Animals , Argentina/epidemiology , Base Sequence , Brain/virology , Chlorocebus aethiops , Genotype , Horse Diseases/epidemiology , Horse Diseases/virology , Horses , Phylogeny , RNA, Viral/chemistry , RNA, Viral/genetics , Sequence Alignment , Sequence Analysis, RNA , Vero Cells , West Nile Fever/epidemiology , West Nile Fever/virology , West Nile virus/classification , West Nile virus/isolation & purificationABSTRACT
Strains of Culex flavivirus (CxFV), an insect virus isolated initially from Japan, were isolated from different species of Culex sp. mosquitoes collected in Corrientes province, Argentina, during 2009. CxFV was detected by reverse transcription polymerase chain reaction and by isolation in C6/36 cell culture. Phylogenetic analysis of nucleotide sequences showed that these strains are related closely to a CxFV strain isolated from Trinidad. Our study represents the first report of CxFV isolation and characterization in Argentina from the same geographic area where West Nile Virus has been detected. Further evaluation and viral competition studies will be necessary to determine the impact of this insect flavivirus on an infection caused by other pathogenic flaviviruses.
Subject(s)
Culex/virology , Flavivirus/isolation & purification , Animals , Argentina , Chlorocebus aethiops , Flavivirus/genetics , Vero CellsABSTRACT
West Nile virus (WNV) was isolated from the brains of 3 horses that died from encephalitis in February 2006. The horses were from different farms in central Argentina and had not traveled outside the country. This is the first isolation of WNV in South America.
Subject(s)
Horse Diseases/virology , West Nile Fever/veterinary , West Nile virus/isolation & purification , Animals , Argentina/epidemiology , Base Sequence/genetics , Brain/virology , Horse Diseases/epidemiology , Horses , Molecular Sequence Data , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction , West Nile Fever/epidemiology , West Nile Fever/virology , West Nile virus/geneticsABSTRACT
Se estudió la secreción de insulina y somatostatina (SRIF) inducida por aloantígenos en el ratón genéticamente diabéticos de la cepa C57BL/KsJ mdb-mdb. Ratones diabéticos (db) inyectados con linfocitos alogeneicos (LA) no mostraron ningún incremento en la segunda fase de secreción estimulada por glucosa 27.5 mM y sólo levemente aumentaron su primera fase. Ratones normales inyectados con LA mostraron un incremento significativo en ambas fases de la secreción de insulina estimulada por glucosa 27.5 mM. La secreción de somatostatina de ratones normales o diabéticos inyectados con linfocitos alogeneicos no difirió de aquella obtenida cuando se inyectaron con linfocitos singeneicos. Linfocitos de ratones db inyectados en ratones alogeneicos causaron un incremento en la secreción de insulina que no difirió significativamente de la producida por la inyección de linfocitos alogeneicos de ratones no diabéticos. Linfocitos de ratón diabético inyectados en ratones singeneicos produjeron una secreción de insulina similar a la producida por la inyección de linfocitos de ratones singeneicos no diabéticos, y menor que la producida por la inyección de linfocitos alogeneicos. En resumen, los ratones db mostraron una respuesta hormonal deteriorada al estímulo antigénico, mientras que mantuvieron su acción aloantigénica
Subject(s)
Mice , Animals , Male , Female , Diabetes Mellitus, Experimental , Insulin/metabolism , Isoantigens/administration & dosage , Lymphocytes/immunology , Somatostatin/metabolismABSTRACT
Se estudió la secreción de insulina y somatostatina (SRIF) inducida por aloantígenos en el ratón genéticamente diabéticos de la cepa C57BL/KsJ mdb-mdb. Ratones diabéticos (db) inyectados con linfocitos alogeneicos (LA) no mostraron ningún incremento en la segunda fase de secreción estimulada por glucosa 27.5 mM y sólo levemente aumentaron su primera fase. Ratones normales inyectados con LA mostraron un incremento significativo en ambas fases de la secreción de insulina estimulada por glucosa 27.5 mM. La secreción de somatostatina de ratones normales o diabéticos inyectados con linfocitos alogeneicos no difirió de aquella obtenida cuando se inyectaron con linfocitos singeneicos. Linfocitos de ratones db inyectados en ratones alogeneicos causaron un incremento en la secreción de insulina que no difirió significativamente de la producida por la inyección de linfocitos alogeneicos de ratones no diabéticos. Linfocitos de ratón diabético inyectados en ratones singeneicos produjeron una secreción de insulina similar a la producida por la inyección de linfocitos de ratones singeneicos no diabéticos, y menor que la producida por la inyección de linfocitos alogeneicos. En resumen, los ratones db mostraron una respuesta hormonal deteriorada al estímulo antigénico, mientras que mantuvieron su acción aloantigénica (AU)