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    • 7. 发明授权
    • Methods using four-layer filter for PCR sample preparation
    • 方法采用四层过滤器进行PCR样品制备
    • US07504363B2
    • 2009-03-17
    • US11016291
    • 2004-12-17
    • Leigh Alexander Burgoyne
    • Leigh Alexander Burgoyne
    • C40B30/00
    • C12N15/1017
    • In accordance with the present invention, there is provided a device, method and kit for detecting any genetic material containing moieties. The present invention enables identification of known or unknown virions or bacteria contained in a fluid therein, such as virions in a blood sample. It also provides for an amenable and highly automatable device for massed screenings and the discovery of sequence variants of known virions, previously undetected virions, and other genetic material containing moieties. In one aspect, the present invention provides a method for storage and analysis of a nucleic acid containing moiety in a biological sample, the method comprising providing a device comprising a plurality of layers, wherein the plurality of layers comprises a collection layer comprising a neutral porous matrix and a dry anticoagulant or liquefying agent, a filter layer comprising a dry solid medium comprising a solid matrix having a particle size limit that precludes eukaryotic cells, but not prokaryotic cells or virions, a separator layer, and an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent; applying a biological sample to the collection layer; filtering remaining components of the biological sample through the collection layer to the filter layer; filtering remaining components of the biological sample through the filter layer to the separator layer; filtering remaining components of the biological sample through the separator layer to the isolation layer; retaining nucleic acid components in the isolation layer while removing a plurality of non-nucleic acid components; drying the isolation layer; providing at least one primer; and analyzing the nucleic acid components using PCR techniques, wherein at least two PCR primers are used.
    • 根据本发明,提供了用于检测含有部分的任何遗传物质的装置,方法和试剂盒。 本发明能够鉴定其中的流体中包含的已知或未知的病毒粒子或细菌,例如血液样品中的病毒粒子。 它还提供了一种适合和高度自动化的装置,用于大规模筛选和发现已知病毒粒子的序列变体,以前未检测到的病毒粒子,以及含有部分的其他遗传物质。 在一个方面,本发明提供了用于存储和分析生物样品中含核酸部分的方法,所述方法包括提供包括多个层的装置,其中所述多个层包括收集层,所述收集层包含中性多孔 基质和干抗凝剂或液化剂,包含干固体培养基的过滤层,所述干固体培养基包含具有排除真核细胞但不包括原核细胞或病毒颗粒的粒度限制的固体基质,隔离层和包含干燥固体的分离层 包含吸收了含有洗涤剂的组合物的中性固体基质的介质; 将生物样品施加到收集层; 通过收集层将生物样品的剩余成分过滤到过滤层; 将生物样品的剩余组分通过过滤层过滤到分离层; 通过隔离层将生物样品的剩余组分过滤到隔离层; 在分离层中保留核酸组分,同时除去多个非核酸组分; 干燥隔离层; 提供至少一个引物; 并使用PCR技术分析核酸成分,其中使用至少两个PCR引物。