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
Y1 - 2001
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.
UR - http://www.scopus.com/inward/record.url?scp=0034888726&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034888726&partnerID=8YFLogxK
U2 - 10.1046/j.1442-200X.2001.01401.x
DO - 10.1046/j.1442-200X.2001.01401.x
M3 - Article
C2 - 11472582
AN - SCOPUS:0034888726
SN - 1328-8067
VL - 43
SP - 372
EP - 378
JO - Pediatrics International
JF - Pediatrics International
IS - 4
ER -