Loc device for genetic analysis with dialysis, chemical lysis, incubation and nucleic acid amplification

Abstract

A lab-on-a-chip (LOC) device for genetic analysis of a biological sample, the LOC device having an inlet for receiving the sample, a supporting substrate, a dialysis section for separating cells larger than a predetermined threshold in the sample from smaller constituents, whereby the cells larger than a predetermined threshold include target cells containing genetic material for analysis, a plurality of reagent reservoirs, a lysis section downstream of the dialysis section for lysing the target cells to release the genetic material therein, the lysis section being in fluid communication with one of the reagent reservoirs containing a lysis reagent for lysing the target cells in the lysis section, an incubation section downstream of the lysis section, the incubation section being in fluid communication with one of the reagent reservoirs containing enzymes for enzymatic reaction with the genetic material, and, a nucleic acid amplification section downstream of the incubation section for amplifying nucleic acid sequences from the genetic material, wherein, the dialysis section, the lysis section, the incubation section and the nucleic acid amplification section are all supported on the supporting substrate.

Description

FIELD OF THE INVENTION[0001]The present invention relates to diagnostic devices that use microsystems technologies (MST). In particular, the invention relates to microfluidic and biochemical processing and analysis for molecular diagnostics.CO-PENDING APPLICATIONS[0002]The following applications have been filed by the Applicant which relate to the present application:GBS001USGBS002USGBS003USGBS005USGBS006USGSR001USGSR002USGAS001USGAS002USGAS003USGAS004USGAS006USGAS007USGAS008USGAS009USGAS010USGAS012USGAS013USGAS014USGAS015USGAS016USGAS017USGAS018USGAS019USGAS020USGAS021USGAS022USGAS023USGAS024USGAS025USGAS026USGAS027USGAS028USGAS030USGAS031USGAS032USGAS033USGAS034USGAS035USGAS036USGAS037USGAS038USGAS039USGAS040USGAS041USGAS042USGAS043USGAS044USGAS045USGAS046USGAS047USGAS048USGAS049USGAS050USGAS054USGAS055USGAS056USGAS057USGAS058USGAS059USGAS060USGAS061USGAS062USGAS063USGAS065USGAS066USGAS067USGAS068USGAS069USGAS070USGAS080USGAS081USGAS082USGAS083USGAS084USGAS085USGAS086USGAS087USGAS...

AI Technical Summary

Benefits of technology

[0095]The lysing process extracts analytical and diagnostic targets from cells in the sample and provides for follow-on processing and analysis of the targets. The lysis subunit being integral to the device, provides for simple assay procedures, low system component-count, and simple system manufacturing procedures, leading into an inexpensive assay system.

[0096]In the incubation section the genetic material undergo various types of preprocessing, like DNA restriction and ligation of adaptor primers, to provide optimal or necessary conditions for the subsequent analytical stages, increasing the informational content of the analytical outcomes and increasing the sensitivity, signal-to-noise-ration, and reliability of the assay system.

[0097]The amplification of target genetic sequences increases the sensitivity and signal-to-noise ratio of the assay system.

[0098]The probe hybridization section provides for analysis of the targets via hybridization. The integrated probe hybridization section provides for an easily usable, mass-producible, and inexpensive integrated solution with low system component-count.

[0099]The integrated image sensor obviates the need for an expensive external imaging system and provides for a mass-producible inexpensive integrated solution with low system component-count that is a compact, light, and highly portable system. The integrated image sensor increases the readout sensitivity by benefiting from large angle of light collection and obviates the need for optical components in the optical collection train.

[0100]The reagent reservoirs, being integral to the LOC device and holding the assay's total reagent requirements, provide for the low system component-count and simple manufacturing procedures, leading into an inexpensive assay system.

Problems solved by technology

Insufficient stringency can result in a high degree of nonspecific binding.

Excessive stringency can lead to a failure of appropriate binding, which results in diminished sensitivity.

Despite the advantages that molecular diagnostic tests offer, the growth of this type of testing in the clinical laboratory has been slower than expected and remains a minor part of the practice of laboratory medicine.

This is primarily due to the complexity and costs associated with nucleic acid testing compared with tests based on methods not involving nucleic acids.

However, controlling fluid flow through the LOC device, adding reagents, controlling reaction conditions and so on necessitate bulky external plumbing and electronics.

Connecting a LOC device to these external devices effectively restricts the use of LOC devices for molecular diagnostics to the laboratory setting.

The cost of the external equipment and complexity of its operation precludes LOC-based molecular diagnostics as a practical option for point-of-care settings.

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