Loc device for pathogen detection with dialysis, thermal lysis and parallel nucleic acid amplification

Abstract

A lab-on-a-chip (LOC) device for detecting pathogens in a biological sample, the LOC device having an inlet for receiving the sample, a supporting substrate, a dialysis section for separating pathogens from larger constituents in the sample, a lysis section downstream of the dialysis section for lysing the pathogens to release genetic material therein, the lysis section having a lysis chamber and a heater for lysing the pathogens while the sample is in the lysis chamber, a first nucleic acid amplification section downstream of the lysis section for amplifying nucleic acid sequences in the genetic material in a first portion of the sample flow from the lysis section, and, a second nucleic acid amplification section downstream of the lysis section for amplifying nucleic acid sequences in the genetic material in a second portion of the sample flow from the lysis section, wherein, the dialysis section, the lysis section, the first nucleic acid amplification section and the second 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

[0102]The easily usable, mass-producible, and inexpensive pathogen detection LOC device accepts a biological sample through its sample receptacle, uses its dialysis section to separate any pathogens contained in the sample, lyses the pathogens as required in its thermal lysis chamber to release the pathogens' genetic materials, amplifies any target genetic sequences, and analyzes the sample's nucleic acid sequences via hybridization with oligonucleotide probes with sensing via its integral imaging array, utilizing reagents stored in the LOC device's reagent reservoirs.

[0108]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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