Kim O, Kim S S
USDA-ARS, ADRU, 319 Bustad, WSU, Pullman, WA 99164, USA.
Acta Virol. 2003;47(2):87-90.
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.
聚合酶链反应（PCR）是检测人类疱疹病毒8型（HHV-8）的一项强大技术，但灵敏度和特异性有限。基于PCR与斑点杂交（DBH）相结合开发了一种新的HHV-8检测方法，并对其灵敏度和特异性进行了评估。使用一对HHV-8特异性引物ORF26out扩增靶DNA。当通过肉眼检测PCR产物时，检测限为从感染HHV-8的BC-3细胞中分离的0.1 ng DNA。对于DBH，用对HHV-8特异的引物对ORF26in扩增的DNA用地高辛（DIG）标记。这种DIG标记的探针能够检测从感染HHV-8的BC-3细胞中分离的1.0 ng DNA。另一方面，PCR与DBH相结合（PCR/DBH）比单独的PCR或DBH更灵敏，而且特异性也很强。PCR/DBH的灵敏度高于单独的PCR和DBH。PCR/DBH检测方法可有效地用于确认临床样本中HHV-8的存在，并特异性地区分HHV-8感染与其他病毒感染。
