Method for identification, selection and analysis of tumour cells

    公开(公告)号:US10895575B2

    公开(公告)日:2021-01-19

    申请号:US14542259

    申请日:2014-11-14

    Abstract: The present invention relates to a method for the diagnosis of tumoural conditions and/or of the corresponding state of advance, wherein a sample from a patient comprising at least one tumour cell is obtained. According to the invention, a purified specimen of the at least one tumour cell is obtained by individually selecting and isolating single cells in a microfluidic device the purified specimen having a purity of at least 90%. On the purified specimen thus obtained there is subsequently performed a molecular analysis such as to highlight a characteristic thereof suited to enabling diagnosis.

    Method and kit for detecting a wild-type and/or a mutated target DNA sequence

    公开(公告)号:US09938574B2

    公开(公告)日:2018-04-10

    申请号:US14439859

    申请日:2013-10-31

    Abstract: The present invention relates to a method for detecting a first and/or a second target DNA sequence from a DNA library, differing in that a mutation generates/eliminates a restriction site for a restriction endonuclease, comprising the steps of: (a) providing the DNA library, in which each of the DNA sequences comprises a first sequence segment, a second sequence segment of genomic DNA as cleaved by the restriction endonuclease, and a third sequence segment reverse complementary to the union of said first sequence segment and 5′ overhang, if any, of the restriction endonuclease; (b) amplifying the library of DNA sequences by PCR using: a first reverse primer which hybridizes to the 3′ end region of the second sequence segment of the first or second target sequence positive strand; a second forward primer which hybridizes to the 3′ end region of the second sequence segment of the first target sequence antipositive strand; a third forward primer comprising a first portion hybridizing to the 5′ end region of the third sequence segment of the second target sequence antipositive strand and a second portion hybridizing to the 3′ end region of the second sequence segment of the second target sequence antipositive strand, wherein the first portion of the third forward primer has a length from 20% to 80% with respect to the total length of the third forward primer; (c) detecting DNA sequences amplified in step (b).

    Method and apparatus for the processing and/or analysis and/or selection of particles, in particular, biological particles
    3.
    发明授权
    Method and apparatus for the processing and/or analysis and/or selection of particles, in particular, biological particles 有权
    用于处理和/或分析和/或选择颗粒,特别是生物颗粒的方法和装置

    公开(公告)号:US09581528B2

    公开(公告)日:2017-02-28

    申请号:US14185648

    申请日:2014-02-20

    Inventor: Nicolò Manaresi

    Abstract: Methods and apparatus are described for the processing (for example washing, incubation, etc.) of particles in which the particles suspended in a first fluid are introduced under laminar flow conditions into at least one first microchamber or first region of the same, in which a second fluid is introduced under laminar flow conditions into at least one second region of the microchamber or of a second microchamber, in such a way as not to mix with the first fluid, and in which at least one field of force acting on the particles is activated in the microchamber(s), to provoke a shift of the particles alone in a predetermined direction and to transfer the same in suspension into the second fluid; an apparatus is preferably used including at least three microchambers n microchambers arranged in sequence with each other in one direction and each connected with the microchamber immediately before it and after it with two orifices offset from each other in a direction perpendicular to the direction of sequence of the microchambers.

    Abstract translation: 描述了用于处理(例如洗涤,孵育等)的颗粒的方法和装置,其中悬浮在第一流体中的颗粒在层流条件下被引入至少一个第一微室或其第一区域中,其中 将第二流体在层流条件下引入至少一个第二区域的微室或第二微室中,以使其不与第一流体混合,并且其中至少一个作用在颗粒上的力场 在微室中激活,以引发颗粒单独在预定方向上的移动并将其悬浮液转移到第二流体中; 优选使用包括在一个方向上彼此依次排列的至少三个微室的至少三个微室,并且每个微孔与紧邻其前的微室相连接,并且之后具有两个孔,所述两个孔在垂直于 微型仓库。

    METHOD AND KIT FOR THE GENERATION OF DNA LIBRARIES FOR MASSIVELY PARALLEL SEQUENCING

    公开(公告)号:US20230250419A1

    公开(公告)日:2023-08-10

    申请号:US17966011

    申请日:2022-10-14

    CPC classification number: C12N15/1082 C12Q1/6806 C12Q1/6855 C12Q1/6869

    Abstract: There is disclosed a method of generating a massively parallel sequencing library comprising the steps of: a) providing a primary WGA DNA library (pWGAlib), including fragments comprising a WGA library universal sequence adaptor; b) re-amplifying the primary WGA DNA library using at least one first primer (1PR) and at least one second primer (2PR); the at least one first primer (1PR) comprising from 5′ to 3′ at least one first sequencing adaptor (1PR5SA), at least one first sequencing barcode (1PR5BC) and a first primer 3′ section (1PR3S) hybridizing to either the WGA library universal sequence adaptor or its reverse complementary; the at least one second primer (2PR) comprising from 5′ to 3′ at least one second sequencing adaptor (2PR5SA) different from the at least one first sequencing adaptor (1PR5SA), and a second primer 3′ section (2PR3S) hybridizing to either the WGA library universal sequence adaptor or its reverse complementary.

    MICROFLUIDIC DEVICE, MICROFLUIDIC SYSTEM AND METHOD FOR THE ISOLATION OF PARTICLES

    公开(公告)号:US20200038870A1

    公开(公告)日:2020-02-06

    申请号:US16342923

    申请日:2017-10-18

    Abstract: A microfluidic device for the isolation of particles of at least one specific type of a sample is described. The microfluidic device comprises: a first inlet adapted to receive a sample comprising the particles of the specific type, at least part of a separation group comprising a separation unit, which comprises a main chamber and a recovery chamber and being adapted to receive the sample and to transfer at least part of the particles of the specific type from the main chamber to the recovery chamber in a selective manner with respect to further particles of the sample, a first outlet which is designed to allow the particles of the specific type to be collected outside of the device and a second outlet adapted to allow at least a portion of the sample to flow out of the main chamber and out of the microfluidic device.

    Method and kit for the generation of DNA libraries for massively parallel sequencing

    公开(公告)号:US11859249B2

    公开(公告)日:2024-01-02

    申请号:US16632268

    申请日:2018-07-20

    CPC classification number: C12Q1/6874 C12N15/1093 C12Q2600/16

    Abstract: There is disclosed a method of generating a massively parallel sequencing library comprising the steps of: a) providing a primary WGA DNA library (pWGAlib), including fragments comprising a WGA library universal sequence adapter; b) performing a single PCR cycle on the pWGAlib using a first primer (1PR) comprising from 5′ to 3′ a first sequencing adapter (1PR5SA) and a first primer 3′ section (1PR3S) hybridizing to the reverse complementary of the WGA library universal sequence adapter; c) performing a single PCR cycle on the on the product of step b) using a second primer (2PR) comprising from 5′ to 3′ a second sequencing adapter (2PR5SA) different from the 1PR5SA, and a second primer 3′ section (2PR3S) hybridizing to the WGA library universal sequence adapter reverse complementary; d) amplifying by PCR the product of step c) using a third primer comprising the 1PR5SA and a fourth primer comprising 2PR5SA.

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