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AssigneeSilicon Biosystems S.P.A.
InventorsNicolò Manaresi, Gianni Medoro, Giuseppe Giorgini
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The present invention relates to a method for the diagnosis of tumoural conditions and/or of the corresponding state of advance, wherein a specimen of an organic fluid taken from the patient and having a high probability of containing at least one circulating tumour cell (CTC) or a disseminated tumour cell (CTD) is enriched in a population of CTCs or CTDs. According to the invention, at least one type of CTCs or CTDs is isolated by selecting individually single cells in a microfluidic device so to constitute a diagnostic specimen having a purity of at least 90%. On the specimen thus obtained there is subsequently performed a molecular analysis such as to highlight a characteristic thereof suited to enabling diagnosis.
A method for the diagnosis of tumoural conditions and/or of the corresponding state of advance, comprising the steps of:
The method according to any one of the preceding claims, characterized in that said microfluidic system isolates said cells in a non-manual way.
The method according to any one of the preceding claims, characterized in that said microfluidic system isolates said cells in an automatic or semi-automatic way.
The method according to any one of the preceding claims, characterized in that said individual selection of said cells is made on the basis of parameters evaluated in the absence of flow of said organic fluid.
The method according to any one of the preceding claims, characterized in that said individual selection of said cells is made on the basis of images.
The method according to the preceding claim, characterized in that said images comprise fluorescence images.
The method according to any one of the preceding claims, characterized in that said individual selection of said cells is made on the basis of impedentiometric properties.
The method according to any one of the preceding claims, characterized in that it comprises a step of labelling of the circulating tumour cells or disseminated tumour cells or portions thereof, with at least one type of tracer.
The method according to the preceding claim, characterized in that said at least one type of tracer is a FISH probe.
The method according to the preceding claim, characterized in that said at least one type of antibody recognizes an antigen chosen in the group consisting of:
The method according to any one of the preceding claims, characterized in that said step of isolating at least one cell or portion of cell from amongst said at least one type of circulating tumour cells or disseminated tumour cells or portions thereof is performed by capturing single cells, preferably each at a specific site of a plurality of sites of said microfluidic device arranged in the microfluidic device according to an array and, subsequently, by selecting single cells from among the captured cells, on the basis of at least one parameter that can be detected by means of a sensor internal or external to the microfluidic device.
The method according to any one of the preceding claims, characterized in that said step of enriching said specimen of said organic fluid comprises a selection of cells made on the basis of at least one parameter chosen in the group consisting of:
The method according to any one of the preceding claims, characterized in that said step of enriching said specimen of said organic fluid in at least one population of cells comprising at least one type of circulating tumour cells or disseminated tumour cells or portions thereof comprises the step of treating said specimen of said organic fluid so as to separate the nucleated cells and enrich it subsequently in nucleated cells.
The method according to the preceding claim, characterized in that said step of enriching said specimen of said organic fluid in at least one population of cells comprising at least one type of circulating tumour cells or disseminated tumour cells or portions thereof comprises at least the step of centrifuging said specimen of said organic fluid in density gradient.
The method according to any one of the preceding claims, characterized in that said step of enriching said specimen of said organic fluid in at least one population of cells comprising at least one type of circulating tumour cells or disseminated tumour cells or portions thereof comprises at least the step of carrying out a positive and/or negative selection via magnetic beads coupled to antibodies according to at least one of the following modalities:
The method according to any one of the preceding claims, characterized in that said at least one enrichment step is performed in combination with said isolation step within the same said microfluidic device used for performing the isolation step.
The method according to the preceding claim, characterized in that a microfluidic device is used equipped with a plurality of different chambers, separated from one another and hydraulically connected, delimited on at least one face by a single chip or by a plurality of separate chips.
The method according to any one of the preceding claims, characterized in that said step of molecular analysis is performed by means of a procedure selected from among:
Dependencies are inferred from the claim text.