Hörling J, Vene S, Franzén C, Niklasson B
Department of Virology, Karolinska Institute, Stockholm, Sweden.
J Clin Microbiol. 1993 Aug;31(8):2004-9. doi: 10.1128/jcm.31.8.2004-2009.1993.
A sensitive assay based on the polymerase chain reaction for the detection of Ockelbo virus RNA was developed. Two primer pairs from the gene coding for the E2 glycoprotein were chosen. By use of a nested strategy for the primers, as few as 1 to 10 PFU could be detected. The amplified products were visualized as bands of appropriate size on ethidium bromide-stained agarose gels. The primer pairs allowed amplification of several Ockelbo and Sindbis virus isolates but discriminated between these and other alphaviruses. Ockelbo virus RNA was detected in 4 of 10 skin biopsy specimens collected during the acute stage of the disease. The identities of the amplified products were confirmed by restriction endonuclease cleavage. Acute- and convalescent-phase sera as well as lymphocytes collected during the convalescent phase were negative by the polymerase chain reaction. No infectious virus could be recovered from any of the specimens.
开发了一种基于聚合酶链反应的灵敏检测方法，用于检测奥克尔博病毒RNA。从编码E2糖蛋白的基因中选择了两对引物。通过对引物采用巢式策略，可检测到低至1至10个空斑形成单位（PFU）。扩增产物在溴化乙锭染色的琼脂糖凝胶上显示为适当大小的条带。这两对引物可扩增几种奥克尔博病毒和辛德毕斯病毒分离株，但能区分这些病毒与其他甲病毒。在疾病急性期采集的10份皮肤活检标本中，有4份检测到奥克尔博病毒RNA。通过限制性内切酶切割确认了扩增产物的身份。急性期和恢复期血清以及恢复期采集的淋巴细胞经聚合酶链反应检测均为阴性。从任何标本中均未分离出感染性病毒。
