Loc device for pathogen detection, genetic analysis and proteomic analysis with dialysis, chemical lysis, incubation and tandem 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 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, an erythrocyte lysis section downstream of the pathogen dialysis section for lysing the erythrocytes with a lysis reagent to release proteins therein, a first leukocyte nucleic acid amplification section downstream of the leukocyte incubation section for amplifying nucleic acid sequences, a second leukocyte nucleic acid amplification section downstream of the first leukocyte nucleic acid amplification section for amplifying nucleic acid sequences, a first pathogen nucleic acid amplification section downstream of the pathogen incubation section for amplifying nucleic acid sequences, and, a second pathogen nucleic acid amplification section downstream of the first pathogen nucleic acid amplification for amplifying nucleic acid sequences, wherein, the leukocyte dialysis section, the pathogen dialysis section, the leukocyte lysis section, the pathogen lysis section, the leukocyte incubation section, the pathogen incubation section, the first and second leukocyte nucleic acid amplification sections and the first and second pathogen nucleic acid amplification sections 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

[0090]The easily usable, mass-producible, and inexpensive LOC device accepts a blood sample, uses a dialysis section for separating the leukocytes and pathogens from the sample, and separately processes the nucleic acid content of the leukocytes and pathogens separated from the blood sample.

[0278]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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