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Acta Virologica Vol.47, p.87-90, 2003 |
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Title: A SENSITIVE AND SPECIFIC DETECTION OF HUMAN HERPESVIRUS 8 BY POLYMERASE CHAIN REACTION AND DOT BLOT HYBRIDIZATION | ||
Author: O. Kim, S.S. Kim | ||
Abstract: Polymerase chain reaction (PCR) is a powerful technique of detecting Human herpesvirus 8 (HHV-8), but has a limited sensitivity and specificity. A new assay of HHV-8 based on combination of PCR with dot blot hybridization (DBH) was developed and evaluated for its sensitivity and specificity. An HHV-8-specific primer pair, ORF26out was used for amplification of target DNA. When the PCR product was detected visually the limit of detection was 0.1 ng DNA isolated from HHV-8-infected BC-3 cells. For DBH, the DNA amplified with the primer pair ORF26in specific for HHV-8 was labeled with digoxigenin (DIG). This DIG-labeled probe was capable of detecting 1.0 ng of DNA isolated from HHV-8-infected BC-3 cells. On the other hand, PCR combined with DBH (PCR/DBH) was more sensitive than PCR or DBH alone and also very specific. The sensitivity of PCR/DBH was higher than that of PCR and DBH alone. The PCR/DBH assay can be applied efficiently to confirm the presence of HHV-8 in clinical samples and to differentiate specifically HHV-8 infection from other viral infections. |
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Keywords: polymerase chain reaction; dot blot hybridization; Human herpesvirus 8 | ||
Year: 2003, Volume: 47, Issue: | Page From: 87, Page To: 90 | |
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