A multiplex PCR assay able to simultaneously detect Torque teno virus isolates from phylogenetic groups 1 to 5
Braz. j. med. biol. res
;
38(6): 853-860, June 2005. ilus, tab
Artículo
en Inglés
| LILACS
| ID: lil-402676
ABSTRACT
Torque teno virus (TTV) is a circular, single-stranded DNA virus that chronically infects healthy individuals of all ages worldwide. TTV has an extreme genetic heterogeneity which is reflected in its current classification into five main phylogenetic groups (1-5). Using specific PCR assays, it has been shown that many individuals are co-infected with TTV isolates belonging to different phylogenetic groups. Here, a multiplex PCR assay was developed, using five recombinant plasmids. Each plasmid carried an insert of different size issued from a TTV isolate belonging to a different group. The assay was able to simultaneously amplify DNAs of TTV isolates belonging to all five phylogenetic groups. Multiplex PCR was then tested satisfactorily on DNAs extracted from 55 serum samples (47 health care workers and 8 AIDS patients). All individuals but nine were infected with at least one TTV isolate. Co-infection with multiple isolates was found in 29/47 (62 percent) health care workers and in 8/8 (100 percent) AIDS patients. A number of discrepancies were observed when results obtained with three thermostable DNA polymerases were compared. For example, four TTV phylogenetic groups were detected in a particular serum sample by using one of the three DNA polymerases, whereas the other two enzymes were able to detect only three TTV groups. However, none of the three enzymes used could be broadly considered to be more efficient than the others. Despite its limitations, the assay described here constitutes a suitable tool to visualize the degree of co-infection of a given population, avoiding time-consuming experiments.
Texto completo:
Disponible
Índice:
LILACS (Américas)
Asunto principal:
Filogenia
/
ADN Viral
/
Reacción en Cadena de la Polimerasa
/
Síndrome de Inmunodeficiencia Adquirida
/
Torque teno virus
/
Infecciones por Virus ADN
Límite:
Humanos
/
Masculino
Idioma:
Inglés
Revista:
Braz. j. med. biol. res
Asunto de la revista:
Biologia
/
Medicina
Año:
2005
Tipo del documento:
Artículo
País de afiliación:
Brasil
Institución/País de afiliación:
Instituto Oswaldo Cruz/BR
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