2001年8月
Quantitation of human herpesvirus 6 DNA in infant with exanthem subitum by microplate PCR-hybridization assay
PEDIATRICS INTERNATIONAL
- ,
- ,
- ,
- ,
- ,
- ,
- ,
- 巻
- 43
- 号
- 4
- 開始ページ
- 372
- 終了ページ
- 378
- 記述言語
- 英語
- 掲載種別
- DOI
- 10.1046/j.1442-200X.2001.01401.x
- 出版者・発行元
- BLACKWELL PUBLISHING ASIA
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 3 1 (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 (154 +/- 76 copies/10(4) PBMC). Furthermore, the virus load in acute phase plasma (53 +/- 75 copies/muL) was also significantly higher than in the convalescent phase samples (2 +/- 9 copies/muL). 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.
Methods: Semiquantitative analysis of the virus genome was carried out in 3 1 (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 (154 +/- 76 copies/10(4) PBMC). Furthermore, the virus load in acute phase plasma (53 +/- 75 copies/muL) was also significantly higher than in the convalescent phase samples (2 +/- 9 copies/muL). 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.
- リンク情報
- ID情報
-
- DOI : 10.1046/j.1442-200X.2001.01401.x
- ISSN : 1328-8067
- PubMed ID : 11472582
- Web of Science ID : WOS:000175879800010