Respiratory syncytial virus (RSV) is significant for public health, capable of causing respiratory tract disease in infants, the elderly and the immunocompromised. The RSV polymerase is an attractive target for antiviral drug development, but as yet, there is no high throughput assay for analyzing RSV polymerase activity, specifically. In this study, using a primer elongation assay as a basis, we analyzed the tolerance of the RSV polymerase for modifications at the 5' end of the primer, and nucleotide analogs. The RSV polymerase was found to accept primers containing 5' biotin or digoxygenin modifications, and nucleotide analogs that are reactive or fluorescent, including 5-ethynyl UTP, 8-azido ATP, 2-amino PTP, and thieno-GTP. These findings provide a menu of options for developing non-isotopic high throughput assays for RSV polymerase RNA synthesis activity, and yield insight regarding the molecular biology of the polymerase complex.

中文翻译：

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呼吸道合胞病毒聚合酶可以用修饰的引物和核苷酸类似物进行RNA合成。

呼吸道合胞病毒（RSV）对于公共卫生非常重要，能够引起婴儿，老年人和免疫功能低下的呼吸道疾病。RSV聚合酶是抗病毒药物开发的有吸引力的靶标，但是，到目前为止，尚无用于分析RSV聚合酶活性的高通量测定法。在这项研究中，我们以引物延伸测定为基础，分析了RSV聚合酶对引物5'末端修饰的耐受性，以及核苷酸类似物。发现RSV聚合酶可以接受含有5'生物素或地高辛配基修饰的引物，以及具有反应性或荧光的核苷酸类似物，包括5-乙炔基UTP，8-叠氮基ATP，2-氨基PTP和硫代GTP。