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

Abstract

A lab-on-a-chip (LOC) device for pathogen detection, genetic analysis and proteomic analysis of a biological sample, the LOC device having an inlet for receiving the sample, a supporting substrate, a leukocyte dialysis section for dividing the sample into a leukocyte stream, and a pathogen and erythrocyte stream containing cells and sample constituents smaller than a first size threshold, a pathogen dialysis section for dividing the pathogen and erythrocyte stream into an erythrocyte stream and a pathogen stream containing cells and sample constituents smaller than a second size threshold, a leukocyte lysis section downstream of the leukocyte dialysis section for lysing the leukocytes with a lysis reagent to release genetic material and proteins therein, a leukocyte incubation section downstream of the leukocyte lysis section for enzymatic reaction of the genetic material with enzymes, a leukocyte nucleic acid amplification section downstream of the leukocyte incubation section for amplifying nucleic acid sequences, a pathogen lysis section downstream of the pathogen dialysis section for lysing the pathogens with a lysis reagent to release genetic material and proteins therein, a pathogen incubation section downstream of the pathogen lysis section for enzymatic reaction of the genetic material with enzymes, a pathogen nucleic acid amplification section downstream of the pathogen incubation section for amplifying nucleic acid sequences, an erythrocyte lysis section downstream of the pathogen dialysis section for lysing the erythrocytes with a lysis reagent to release proteins therein, wherein, the leukocyte dialysis section, the pathogen dialysis section, the leukocyte lysis section, the pathogen lysis section, the erythrocyte lysis section, the leukocyte incubation section, the pathogen incubation section, the leukocyte nucleic acid amplification section and the pathogen 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

[0106]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.

[0107]In the incubation section the genetic material undergo various types of preprocessing, like nucleic acid 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.

[0108]The amplification of target genetic sequences increases the sensitivity and signal-to-noise ratio of the assay system.

[0109]The probe hybridiz

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

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