Genetic analysis loc for nucleic acid amplification using nucleic acid sequence based amplification

Abstract

A lab-on-a-chip (LOC) device for genetic analysis of a sample containing target nucleic acid sequences, the LOC device having a sample inlet for receiving the sample, a plurality of reagent reservoirs containing dNTPs, primers, reverse transcriptase, RNA polymerase, and buffer solution for addition to the sample to form an amplification mixture, and, a nucleic acid amplification section for maintaining the amplification mixture at a predetermined temperature during isothermal amplification of the target nucleic acid sequences.

Description

FIELD OF THE INVENTIONThe 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 APPLICATIONSThe following applications have been filed by the Applicant which relate to the present application:GBS001USGBS002USGBS003USGBS005USGBS006USGSR001USGSR002USGAS001USGAS002USGAS003USGAS004USGAS006USGAS007USGAS008USGAS009USGAS010USGAS012USGAS013USGAS014USGAS015USGAS016USGAS017USGAS018USGAS019USGAS020USGAS021USGAS022USGAS023USGAS024USGAS025USGAS026USGAS027USGAS028USGAS030USGAS031USGAS032USGAS033USGAS034USGAS035USGAS036USGAS037USGAS038USGAS039USGAS040USGAS041USGAS042USGAS043USGAS044USGAS045USGAS046USGAS047USGAS048USGAS049USGAS050USGAS054USGAS055USGAS056USGAS057USGAS058USGAS059USGAS060USGAS061USGAS062USGAS063USGAS065USGAS066USGAS067USGAS068USGAS069USGAS070USGAS080USGAS081USGAS082USGAS083USGAS084USGAS085USGAS086USGAS087USGAS088USGAS089U...

AI Technical Summary

Benefits of technology

This LOC device has the advantage that thermal cycling is not required, which simplifies thermal control electronics, allows more uniform temperature control, and reduces degradation of materials in the LOC device. This LOC device design will reduce the degree of evaporation during operation, allowing improved control over the physical and chemical conditions within the LOC device. This LOC device has the advantages provided by sequence-specific amplification, including: sensitivity provided by amplification; broad dynamic range; and high specificity for the target RNA sequence.

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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