RESUMEN
The FTA card has emerged as a promising alternative for nucleic acid extraction. The FTA card is a filter paper impregnated with chemicals that preserve and stabilize the genetic material present in the sample, allowing for its storage and transport at room temperature. The aim of this study was to test the card for the detection of RNA and DNA nucleic acids. Two RNA viruses (Senecavirus A and classical swine fever virus) and two DNA viruses (African swine fever virus and suid alphaherpesvirus 1) were tested, and in all cases, there was a decrease in sensitivity. The methods exhibited good repeatability and demonstrated a rapid and practical use for sample transport and nucleic acid extraction.
Asunto(s)
Virus de la Fiebre Porcina Africana , Animales , Porcinos , Virus de la Fiebre Porcina Africana/aislamiento & purificación , Virus de la Fiebre Porcina Africana/genética , Virus de la Fiebre Porcina Clásica/genética , Virus de la Fiebre Porcina Clásica/aislamiento & purificación , Herpesvirus Suido 1/aislamiento & purificación , Herpesvirus Suido 1/genética , ARN Viral/genética , ARN Viral/aislamiento & purificación , Medicina Veterinaria/métodos , Enfermedades de los Porcinos/virología , Enfermedades de los Porcinos/diagnóstico , Virus ADN/genética , Virus ADN/aislamiento & purificación , Picornaviridae/genética , Picornaviridae/aislamiento & purificación , Picornaviridae/clasificación , Sensibilidad y Especificidad , ADN Viral/genética , Virus ARN/genética , Virus ARN/aislamiento & purificación , Virus ARN/clasificación , Infecciones por Virus ADN/veterinaria , Infecciones por Virus ADN/diagnóstico , Infecciones por Virus ADN/virología , Manejo de Especímenes/métodos , Manejo de Especímenes/instrumentaciónRESUMEN
INTRODUCTION: Torque teno virus (TTV) and SEN virus are circular single-stranded DNA viruses that cause blood-borne infections. The SEN virus (SEN-V) was originally detected in the serum of an injection drug user infected with human immunodeficiency virus (HIV). Recently TTV was discovered as a potential causative agent of non-A-E hepatitis. The aim of this study was to investigate the prevalence of the SEN-V-D/H and TTV in HIV patients and healthy blood donors in Iran. METHODS: One hundred and fifty HIV patients with a mean age of 50.46 ± 18.46 years and 150 healthy blood donors with a mean age of 48.16 ± 13.73 years were included in this study. TTV and SEN-V were detected by the PCR and were quantitatively assayed by competitive PCR (nested and semi-nested PCR). Restriction fragment length polymorphisms (RFLPs) were used to determine the heterogeneity of TTV. RESULTS: TTV and SEN-V were detected 96 (64%) and 84 (56%) of 150 HIV patients respectively. These rates were 34% (n=51) and 37.33% (n=56) in healthy blood donors (significant, p<0.05). PCR detected SEN-V/TTV DNA from 32 of the healthy blood donors (21.33%), while 65 (43.33%) of HIV patients were positive for SEN-V/TTV DNA. Of 150 HIV patients, 32.66% and 23.33% were positive for SEN-V-H and SEN-V-D, respectively and 18.66% (n=28) were co-infected with SEN-V-D/H. CONCLUSIONS: The prevalence of SEN-VD/H and TTV is higher in HIV patients than in healthy blood donors in Southern Iran. Our results suggest that TTV and SEN-V might play a role in the development of liver disease in patients with immunodeficiency diseases.
Asunto(s)
Infecciones por Virus ADN/virología , Infecciones por VIH/virología , Torque teno virus/genética , Adulto , Anciano , Donantes de Sangre , Coinfección/virología , Infecciones por Virus ADN/diagnóstico , Infecciones por Virus ADN/epidemiología , ADN Viral/análisis , Femenino , Genotipo , Infecciones por VIH/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
Introduction Torque teno virus (TTV) and SEN virus are circular single-stranded DNA viruses that cause blood-borne infections. The SEN virus (SEN-V) was originally detected in the serum of an injection drug user infected with human immunodeficiency virus (HIV). Recently TTV was discovered as a potential causative agent of non-A-E hepatitis. The aim of this study was to investigate the prevalence of the SEN-V-D/H and TTV in HIV patients and healthy blood donors in Iran. Methods One hundred and fifty HIV patients with a mean age of 50.46 ± 18.46 years and 150 healthy blood donors with a mean age of 48.16 ± 13.73 years were included in this study. TTV and SEN-V were detected by the PCR and were quantitatively assayed by competitive PCR (nested and semi-nested PCR). Restriction fragment length polymorphisms (RFLPs) were used to determine the heterogeneity of TTV. Results TTV and SEN-V were detected 96 (64%) and 84 (56%) of 150 HIV patients respectively. These rates were 34% (n=51) and 37.33% (n=56) in healthy blood donors (significant, p<0.05). PCR detected SEN-V/TTV DNA from 32 of the healthy blood donors (21.33%), while 65 (43.33%) of HIV patients were positive for SEN-V/TTV DNA. Of 150 HIV patients, 32.66% and 23.33% were positive for SEN-V-H and SEN-V-D, respectively and 18.66% (n=28) were co-infected with SEN-V-D/H. Conclusions The prevalence of SEN-VD/H and TTV is higher in HIV patients than in healthy blood donors in Southern Iran. Our results suggest that TTV and SEN-V might play a role in the development of liver disease in patients with immunodeficiency diseases. .
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Infecciones por Virus ADN/virología , Infecciones por VIH/virología , Torque teno virus/genética , Donantes de Sangre , Coinfección/virología , Infecciones por Virus ADN/diagnóstico , Infecciones por Virus ADN/epidemiología , ADN Viral/análisis , Genotipo , Infecciones por VIH/epidemiología , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
This study investigates the frequency of Torque teno virus (TTV) infection in 150 blood donors and 77 patients requiring haemodialysis in southern Brazil. Plasma samples were screened for TTV DNA using polymerase chain reaction (PCR). The prevalences of TTV among blood donors and patients requiring haemodialysis were 73.3% and 68.8%, respectively. The presence of TTV was correlated with age in the blood donors (p = 0.024). In haemodialysis patients, no association was found between TTV infection and the demographic parameters (age, sex and education), the duration of haemodialysis or a history of blood transfusion. This study is the first to evaluate the prevalence of TTV infection in Brazilian patients requiring haemodialysis.
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Donantes de Sangre , Infecciones por Virus ADN/epidemiología , Diálisis Renal , Torque teno virus/aislamiento & purificación , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Brasil/epidemiología , Estudios Transversales , Infecciones por Virus ADN/sangre , Infecciones por Virus ADN/diagnóstico , Escolaridad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Prevalencia , Factores de Riesgo , Factores de Tiempo , Torque teno virus/genética , Adulto JovenRESUMEN
This study investigates the frequency of Torque teno virus (TTV) infection in 150 blood donors and 77 patients requiring haemodialysis in southern Brazil. Plasma samples were screened for TTV DNA using polymerase chain reaction (PCR). The prevalences of TTV among blood donors and patients requiring haemodialysis were 73.3% and 68.8%, respectively. The presence of TTV was correlated with age in the blood donors (p = 0.024). In haemodialysis patients, no association was found between TTV infection and the demographic parameters (age, sex and education), the duration of haemodialysis or a history of blood transfusion. This study is the first to evaluate the prevalence of TTV infection in Brazilian patients requiring haemodialysis.
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Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Donantes de Sangre , Infecciones por Virus ADN/epidemiología , Diálisis Renal , Torque teno virus/aislamiento & purificación , Brasil/epidemiología , Estudios Transversales , Infecciones por Virus ADN/sangre , Infecciones por Virus ADN/diagnóstico , Escolaridad , Reacción en Cadena de la Polimerasa , Prevalencia , Factores de Riesgo , Factores de Tiempo , Torque teno virus/genéticaRESUMEN
In this study, 40 pigs with respiratory and wasting disorders from Cuban swine herds were screened by PCR for the presence of TTSuV1, TTSuV2, PCV-2, PPV and CSFV in spleen samples. The variability of the porcine TTSuV sequences obtained was investigated by phylogenetic analysis. This study showed for the first time that TTSuV1 and TTSuV2 were present in Cuban swine herds. The investigation revealed the following infection rates: TTSuV1 40%, TTSuV2 37.5%, PCV-2 70%, PPV 37.5% and CSFV in 52.5%. The presence of two or more of these viruses at different rates in the same spleen samples was revealed. Also, a higher genetic diversity of TTSuV2 sequences was observed regarding TTSuV1 sequences.
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Infecciones por Virus ADN/veterinaria , Bazo/virología , Enfermedades de los Porcinos/virología , Torque teno virus/clasificación , Torque teno virus/aislamiento & purificación , Animales , Secuencia de Bases , Circovirus/aislamiento & purificación , Virus de la Fiebre Porcina Clásica/aislamiento & purificación , Infecciones por Virus ADN/diagnóstico , Infecciones por Virus ADN/virología , ADN Viral/genética , Parvovirus Porcino/aislamiento & purificación , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , Porcinos , Enfermedades de los Porcinos/diagnóstico , Torque teno virus/genéticaRESUMEN
Oysters Crassostrea gigas were placed at water supply canals of three shrimp farms in Guasave, Mexico where WSSV outbreaks occur. Animals were sampled through April-August and September-December to detect WSSV DNA. By using three different PCR protocols, only oysters from a farm undergoing a WSSV outbreak were found WSSV-positive in gills and digestive gland. Two WSSV amplicons were sequenced and they corresponded over 99% to WSSV genome segments. Results showed that oysters can capture WSSV particles suspended in water. Susceptibility of oysters to WSSV infection and their role as a carrier remain to be determined.
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Crassostrea/virología , Infecciones por Virus ADN/veterinaria , Mariscos/virología , Virus del Síndrome de la Mancha Blanca 1/aislamiento & purificación , Animales , Portador Sano/veterinaria , Portador Sano/virología , Infecciones por Virus ADN/diagnóstico , Infecciones por Virus ADN/transmisión , ADN Viral/genética , Sistema Digestivo/virología , Branquias/virología , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico , Microbiología del Agua , Abastecimiento de Agua/análisisRESUMEN
The prevalence of TT virus (TTV) infection was investigated by Polymerase Chain Reaction (PCR) in low- (blood donors and healthy children/adolescents) and high-risk (hemophiliacs) groups from São Paulo, Brazil. Primers based on the untranslated region (UTR) of the viral genome proved to be much more ubiquitous, leading to much higher frequencies for both groups (>or= 81%) than the earlier N22-PCR directed to the open reading frame 1 (blood donors, 5.5%, and hemophiliacs, 42.3%). The UTR-PCR also revealed an interesting profile for healthy children/adolescents: very high prevalence at the early years and significant decrease in male teenagers. The N22-PCR, in turn, demonstrated higher frequency in hemophiliacs treated with fresh blood products (58%), than in those treated with virus-inactivated clotting factors (9.4%) and blood donors (5.5%).
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Infecciones por Virus ADN/epidemiología , Torque teno virus , Adolescente , Donantes de Sangre , Brasil/epidemiología , Niño , Preescolar , Infecciones por Virus ADN/diagnóstico , Femenino , Hemofilia A/virología , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Prevalencia , Estudios Seroepidemiológicos , Regiones no TraducidasRESUMEN
Recombinant plasmids carrying 199 base pairs (bp) inserts from the non coding region (nucleotides (nt) 6-204) of the TT virus (TTV) genome were used to standardize an heteroduplex mobility assay able to detect mixed infections of a single individual with several TTV isolates. In this simplified heteroduplex mobility assay, polymerase chain reaction (PCR) products were analyzed directly by polyacrylamide gel electrophoresis, without requirement for post-PCR denaturation and annealing steps of the amplicons. The assay was used to test TTV positive serum samples collected from healthy 1-7 years old children, 11-17 years old adolescents, and 24-39 years old blood donors living in Rio de Janeiro, Brazil, as well as TTV positive samples from Amazonian Indians. The results showed a very high frequency of multiple infection in all groups, with 20/30 (67%), 31/33 (94%), 35/38 (92%), and 34/37 (92%) of the samples collected from children, adolescents, blood donors, and Amazonian Indians, respectively, containing more than one TTV genotype.
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Infecciones por Virus ADN/diagnóstico , ADN Viral/análisis , Análisis Heterodúplex/métodos , Torque teno virus/aislamiento & purificación , Adolescente , Adulto , Disparidad de Par Base , Donantes de Sangre , Brasil/epidemiología , Niño , Preescolar , Infecciones por Virus ADN/epidemiología , ADN Viral/sangre , Femenino , Genotipo , Estado de Salud , Humanos , Indígenas Sudamericanos , Lactante , Masculino , Reacción en Cadena de la Polimerasa , Prevalencia , Estudios Seroepidemiológicos , Torque teno virus/genética , Población UrbanaRESUMEN
TT virus (TTV) is a newly described nonenveloped human virus, with a circular, negative-stranded DNA genome, that was first identified in the blood of a patient with posttransfusion hepatitis of unknown etiology. PCR primers and conditions used for TTV DNA amplification may greatly influence the level of TTV detection in serum. Three PCR assays, with different regions of the genome as targets, were used to test TTV DNA in 130 sera from children and adults visiting a hospital in the south of Brazil, most of them for routine procedure. Forty-four percent of adult sera and 73% of sera from children aged 0-10 years were TTV positive with at least one PCR assay. However, the three assays were able to detect only 33%, 35%, and 70% of the total positive samples. Our results showed a high prevalence of TTV infection in the south of Brazil, particularly among young children, and confirmed the necessity of performing several PCR assays to assess the true TTV prevalence in a determined population.
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Infecciones por Virus ADN/epidemiología , Reacción en Cadena de la Polimerasa/métodos , Torque teno virus/aislamiento & purificación , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Brasil/epidemiología , Niño , Preescolar , Cartilla de ADN , Infecciones por Virus ADN/sangre , Infecciones por Virus ADN/diagnóstico , ADN Viral/sangre , ADN Viral/aislamiento & purificación , Pruebas Diagnósticas de Rutina , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Técnicas de Amplificación de Ácido Nucleico/métodos , Prevalencia , Estudios Seroepidemiológicos , Torque teno virus/genéticaRESUMEN
TT virus (TTV) is a newly described nonenveloped human virus, with a circular, negative-stranded DNA genome, that was first identified in the blood of a patient with posttransfusion hepatitis of unknown etiology. PCR primers and conditions used for TTV DNA amplification may greatly influence the level of TTV detection in serum. Three PCR assays, with different regions of the genome as targets, were used to test TTV DNA in 130 sera from children and adults visiting a hospital in the south of Brazil, most of them for routine procedure. Forty-four percent of adult sera and 73 percent of sera from children aged 0-10 years were TTV positive with at least one PCR assay. However, the three assays were able to detect only 33 percent, 35 percent, and 70 percent of the total positive samples. Our results showed a high prevalence of TTV infection in the south of Brazil, particularly among young children, and confirmed the necessity of performing several PCR assays to assess the true TTV prevalence in a determined population