Quantitation of human herpesvirus 6 DNA in infant with exanthem subitum by microplate PRC-hybridization assay

Tomoko Abe, Tetsushi Yoshikawa, Masaru Ihira, Kyoko Suzuki, Sadao Suga, Mikio Nishida, Minoru Nagata, Yoshizo Asano

Research output: Contribution to journalArticle

4 Citations (Scopus)

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/104 PBMC) was significantly higher than during the convalescent phase (54±76 copies/104 PBMC). Furthermore, the virus load in acute phase plasma (53 ± 75 copies/μL) was also significantly higher than in the convalescent phase samples (2 ± 9 copies/μL). 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.

Original languageEnglish
Pages (from-to)372-378
Number of pages7
JournalPediatrics International
Volume43
Issue number4
DOIs
Publication statusPublished - 03-09-2001

Fingerprint

Human Herpesvirus 6
Exanthema
Viruses
DNA
Blood Cells
Genome
Herpesviridae Infections
Enzyme-Linked Immunosorbent Assay
Polymerase Chain Reaction
Human Herpesvirus 7
Sick Leave
Virus Diseases
Cytomegalovirus
Limit of Detection
Cell Count

All Science Journal Classification (ASJC) codes

  • Pediatrics, Perinatology, and Child Health

Cite this

Abe, Tomoko ; Yoshikawa, Tetsushi ; Ihira, Masaru ; Suzuki, Kyoko ; Suga, Sadao ; Nishida, Mikio ; Nagata, Minoru ; Asano, Yoshizo. / Quantitation of human herpesvirus 6 DNA in infant with exanthem subitum by microplate PRC-hybridization assay. In: Pediatrics International. 2001 ; Vol. 43, No. 4. pp. 372-378.
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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/104 PBMC) was significantly higher than during the convalescent phase (54±76 copies/104 PBMC). Furthermore, the virus load in acute phase plasma (53 ± 75 copies/μL) was also significantly higher than in the convalescent phase samples (2 ± 9 copies/μL). 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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Quantitation of human herpesvirus 6 DNA in infant with exanthem subitum by microplate PRC-hybridization assay. / Abe, Tomoko; Yoshikawa, Tetsushi; Ihira, Masaru; Suzuki, Kyoko; Suga, Sadao; Nishida, Mikio; Nagata, Minoru; Asano, Yoshizo.

In: Pediatrics International, Vol. 43, No. 4, 03.09.2001, p. 372-378.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Quantitation of human herpesvirus 6 DNA in infant with exanthem subitum by microplate PRC-hybridization assay

AU - Abe, Tomoko

AU - Yoshikawa, Tetsushi

AU - Ihira, Masaru

AU - Suzuki, Kyoko

AU - Suga, Sadao

AU - Nishida, Mikio

AU - Nagata, Minoru

AU - Asano, Yoshizo

PY - 2001/9/3

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N2 - 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/104 PBMC) was significantly higher than during the convalescent phase (54±76 copies/104 PBMC). Furthermore, the virus load in acute phase plasma (53 ± 75 copies/μL) was also significantly higher than in the convalescent phase samples (2 ± 9 copies/μL). 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.

AB - 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/104 PBMC) was significantly higher than during the convalescent phase (54±76 copies/104 PBMC). Furthermore, the virus load in acute phase plasma (53 ± 75 copies/μL) was also significantly higher than in the convalescent phase samples (2 ± 9 copies/μL). 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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