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首页> 外文期刊>Journal of Virological Methods >Single-tube fluorescent product-enhanced reverse transcriptase assay with Ampliwax (TM) (STF-PERT) for retrovirus quantitation
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Single-tube fluorescent product-enhanced reverse transcriptase assay with Ampliwax (TM) (STF-PERT) for retrovirus quantitation

机译:使用Ampliwax(TM)(STF-PERT)进行单管荧光产物增强的逆转录酶测定以进行逆转录病毒定量

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摘要

A TaqMan fluorescent probe-based product enhanced reverse transcriptase (RT) assay is described in which the RT and polymerase chain reaction (PCR) steps are set-up in a single tube, in two compartments separated by Ampliwax(TM) (designated as single-tube fluorescent product-enhanced reverse transcriptase assay (STF-PERT)). This simplification of the two-step method resulted in increased assay reproducibility and handling efficiency while maintaining the sensitivity of the PERT assay ( < 10 virions). The STF-PERT assay can be used to quantitate low amounts of retrovirus in clinical and research materials and to evaluate retrovirus contamination in cell substrates and biological products in human use.
机译:描述了一种基于TaqMan荧光探针的产品增强逆转录酶(RT)分析方法,其中RT和聚合酶链反应(PCR)步骤设置在单个试管中,并在两个由Ampliwax(TM)分隔的隔室中(指定为单个管荧光产品增强的逆转录酶测定法(STF-PERT))。两步方法的这种简化导致了测定的可重复性和处理效率的提高,同时保持了PERT测定的灵敏度(<10病毒体)。 STF-PERT分析可用于定量临床和研究材料中的少量逆转录病毒,并评估人类使用的细胞基质和生物产品中的逆转录病毒污染。

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