Quantitation of human herpesvirus 6 DNA in infant with exanthem subitum by microplate PCR-hybridization assay.
Pediatr Int
; 43(4): 372-8, 2001 Aug.
Article
en En
| MEDLINE
| ID: mdl-11472582
ABSTRACT
BACKGROUND:
Quantitative analysis of human herpesvirus 6 (HHV-6) genome is important for monitoring active virus infection. The purpose of our study is to evaluate the reliability of a hybridization-based microtiter plate assay (polymerase chain reaction enzyme-linked immunosorbent assay (PCR ELISA)) for quantifying the virus genome.METHODS:
Semiquantitative analysis of the virus genome was carried out in 31 (18 male and 13 female) infants with primary HHV-6 infection. If the HHV-6 virus could be isolated from the peripheral blood mononuclear cells (PBMC), the infants were defined as being infected with HHV-6. The PCR ELISA method was used to determine the virus load. A titration of the virus was also carried out in the samples obtained during the acute phase of exanthem subitum.RESULTS:
Specificity of the method was demonstrated by a lack of amplification of human herpesvirus 7 and cytomegalovirus DNA. The upper and lower detection limits of the method were 58 and 5800 copies of the virus genome, respectively. The quantity of HHV-6 DNA in the PBMC during the acute phase (879 +/- 975 copies/10(4) PBMC) was significantly higher than during the convalescent phase (54 +/- 76 copies/10(4) PBMC). Furthermore, the virus load in acute phase plasma (53 +/- 75 copies/microL) was also significantly higher than in the convalescent phase samples (2 +/- 9 copies/microL). Virus load in both PBMC and plasma gradually increased after the onset of exanthem subitum until about day 3 to 4 of the illness, but then decreased quickly. However, there was no significant association between virus load and the numbers of infected cells.CONCLUSION:
Virus load in both PBMC and plasma gradually increased after the onset of exanthem subitum until about day 3 and day 4 of the illness, respectively, then it decreased quickly. These results indicate that our PCR ELISA system is reliable for monitoring active HHV-6 infection in vivo.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
ADN Viral
/
Ensayo de Inmunoadsorción Enzimática
/
Reacción en Cadena de la Polimerasa
/
Herpesvirus Humano 6
/
Exantema Súbito
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
Límite:
Female
/
Humans
/
Infant
/
Male
Idioma:
En
Revista:
Pediatr Int
Asunto de la revista:
PEDIATRIA
Año:
2001
Tipo del documento:
Article
País de afiliación:
Japón