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Anal. Chem:微生物所在微滴数字PCR技术研制方面取得进展

2019-01-08 MedSci 微生物研究所

数字PCR是一种用于绝对核酸定量检测方法,具有前所未有的准确度和精确度,是继实时荧光定量PCR技术之后的第三代核酸定量扩增检测技术,在液体活检、肿瘤伴随诊断、无创产前筛查、病原载量监测等方面具有重要应用前景,是科研和临床领域的平台级新技术。数字PCR市场主流产品主要为欧美跨国公司的高端仪器,在科研和临床的应用还比较初步。数字PCR的更广泛使用和临床应用,仍然存在几个主要挑战,包括降低成本,集成仪器

数字PCR是一种用于绝对核酸定量检测方法,具有前所未有的准确度和精确度,是继实时荧光定量PCR技术之后的第三代核酸定量扩增检测技术,在液体活检、肿瘤伴随诊断、无创产前筛查、病原载量监测等方面具有重要应用前景,是科研临床领域的平台级新技术。数字PCR市场主流产品主要为欧美跨国公司的高端仪器,在科研临床的应用还比较初步。数字PCR的更广泛使用和临床应用,仍然存在几个主要挑战,包括降低成本,集成仪器平台和简化实验操作。

中国科学院微生物研究所微生物资源前期开发国家重点实验室杜文斌团队最近报道了一种可重复使用的微流体芯片,该芯片基于阶梯乳化(Step Emulsification)产生纳升液滴阵列,用于同时对8个样品进行片上多重荧光数字PCR。装置包含两块玻璃板,可实现预先填充矿物油的快速组装。通过由单个压力泵同时驱动8个样品通道的阶梯乳化,利用喷嘴阵列快速产生液滴,生成的液滴在U形腔室中可自组装成单层液滴阵列,每个腔室可容纳约10000个0.35 nL体积的微滴。系统采用的油相配方有效避免和消除了数字PCR温控过程中气泡的产生,在热循环期间不需要采用过压控制。团队将该芯片应用于临床样本HER2拷贝数变异的定量评估,验证了多重数字PCR性能,平行检测结果与美国赛默飞公司的QuantStudio 3D数字PCR仪一致,这对于乳腺癌的靶向治疗和预后具有重要临床应用价值。这种低成本、可重复使用且用户友好的设备可广泛应用于各种应用,在普通实验室仅需一台平板PCR仪和小型气压泵即可开展数字PCR扩增。芯片可在拆卸和简单清洁程序后重复使用50次以上,显着降低了单次数字PCR检测的耗材成本。

上述研究结果于2019年1月4日在线发表于国际学术期刊Analytical Chemistry。

原始出处:

Mengyue Nie,et al.Assembled Step Emulsification Device for Multiplex Droplet Digital PCR.Anal. Chem. January 4, 2019

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    2019-07-11 AspirantSuo
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    2019-10-07 一个字-牛

    学习了谢谢分享

    0

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    学习了谢谢分享

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    2019-01-28 hb2008ye
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    2019-01-10 Tommy1950
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    2019-01-08 1209e435m98(暂无昵称)

    学习了,谢谢分享

    0

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