Campbell W P, Huang C
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509, USA.
J Virol Methods. 1995 May;53(1):55-61. doi: 10.1016/0166-0934(94)00176-h.
Universal primers have been identified and a protocol developed that are suitable for rapid detection of California encephalitis (CE) complex viruses in a reverse transcription-polymerase chain reaction (RT-PCR) assay. These primers correspond to sequences in the coding regions of the G2 glycoprotein of the middle-size RNA segment. The identities of the amplified products were confirmed by sequencing on the clones or PCR products. The technique is capable of detecting 40 plaque-forming units (PFU) directly on an ethidium bromide-stained agarose gel and the sensitivity increases to 0.4-1 PFU when a radiolabeled probe is used as the detector.
已经鉴定出通用引物并开发了一种方案,适用于在逆转录-聚合酶链反应(RT-PCR)分析中快速检测加利福尼亚脑炎(CE)复合病毒。这些引物对应于中号RNA片段G2糖蛋白编码区的序列。通过对克隆或PCR产物进行测序来确认扩增产物的身份。该技术能够直接在溴化乙锭染色的琼脂糖凝胶上检测40个噬斑形成单位(PFU),当使用放射性标记探针作为检测器时,灵敏度可提高到0.4-1 PFU。