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[目的]利用呼吸道合胞病毒(RSV)A、B亚型特异性cDNA探针,检测和鉴别病毒分离株或临床标本,分析广州地区分离株RSV亚型。[方法]利用RSV两个亚型病毒C蛋白胞外区段编码基因的cDNA克隆片段,随机引物法标记,合成地高辛甙元标记RSV亚型特异的探针,检测12株广州地区分离株和120份临床标本。[结果]两型探针均可检出1pg由RSV纯病毒颗粒提取的RNA,两亚型间无交叉反应;12株分离株全部为A亚型,120份临床标本检出9例A亚型与病毒分离结果一致,未检出B亚型病毒。[结论]用非同位毒标记RSV亚型特异性探针,具有很高的特异性和敏感性,其操作快速、简便、安全,可在临床应用。临床初步应用结果表明,广州地区临床分离株以A亚型为主。
[Objective] To detect and identify virus isolates or clinical samples using RSV A and B subtype-specific cDNA probes to analyze RSV subtypes of Guangzhou isolates. [Method] The cloned fragment of gene encoding extracellular domain of C protein of two subtypes of RSV was screened by random primers and labeled with digoxigenin. RSV isotype specific probes were detected in 12 strains of Guangzhou isolates and 120 Clinical specimens. [Results] The RNA extracted from 1 pg of RSV virion could be detected by both probes. There was no cross-reaction between the two subtypes. All 12 isolates were subtypes A, and 120 subtypes were found in 120 clinical samples Consistent with the results of virus isolation, no subtype B virus was detected. [Conclusion] It is of high specificity and sensitivity to label RSV subtype specific probe by non-isotopic virus. The method is rapid, simple and safe and can be used clinically. Clinical preliminary application results show that the clinical isolates in Guangzhou to A subtype.