Toward a Rapid, Integrated, and Fully Automated DNA Diagnostic Assay for Chlamydia trachomatis and Neisseria gonorrhoeae

Pourahmadi, F.; Taylor, Mike; Kovacs, Greg; Lloyd, Kristen; Sakai, Stan; Schafer, Tamlyn; Helton, Bret; Western, Linda M.; Zaner, Sandy; Ching, Jesus; McMillan, Bill; Belgrader, Phil; Northrup, M. Allen

doi:10.1093/clinchem/46.9.1511

Cited by 13 publications

(3 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…13,15,19 Clinical applications for these devices can be separated into different categories based on their target analytes and the method of analysis. These categories may include immunoassays, [46][47][48] proteins, peptides, and amino acids, [49][50][51][52][53] genetic material such as DNA or RNA, 54,55 metabolites, therapeutic drugs and other small molecules, 56,57 and cellular assays 11,[58][59][60] to mention the major applications.…”

Section: Recent Progress In Clinical Applicationsmentioning

confidence: 99%

“…110 Most intracellular component analyses employed PCR methods. 11,[58][59][60]111 Belgrader et al reported several new designs demonstrating rapid detection and identification of spores and characterization of a SNP for the hereditary hemochromatosis gene. 58 Similar to Belgrader's research, Yuen et al constructed a computer numerical control-machined plexiglass-based microchip to isolate white blood cells from a small volume of human whole blood, as part of sample preparation for the following PCR reactions.…”

Section: Cell Analysismentioning

confidence: 99%

See 1 more Smart Citation

Recent progress in the development of μTAS for clinical analysis

Yan

Garcı́a

Henry

2003

Analyst

View full text Add to dashboard Cite

show abstract

Section: Recent Progress In Clinical Applicationsmentioning

confidence: 99%

Section: Cell Analysismentioning

confidence: 99%

Recent progress in the development of μTAS for clinical analysis

Yan

Garcı́a

Henry

2003

Analyst

View full text Add to dashboard Cite

show abstract

“…Over the past 10 years this drive towards integration has taken two main approaches. The first option is integration of PCR with downstream analysis, typically in the form of a microarray, 25 quantitative PCR (qPCR), [26][27][28] or more commonly CE. 29,30 The second perspective has been to integrate PCR amplification with upstream SP.…”

Section: Introductionmentioning

confidence: 99%

Optimization of multiplexed PCR on an integrated microfluidic forensic platform for rapid DNA analysis

Estes

Yang

Duane

et al. 2012

Analyst

View full text Add to dashboard Cite

This study reports the design, prototyping, and assay development of multiplexed polymerase chain reaction (PCR) on a plastic microfluidic device. Amplification of 17 DNA loci is carried out directly on-chip as part of a system for continuous workflow processing from sample preparation (SP) to capillary electrophoresis (CE). For enhanced performance of on-chip PCR amplification, improved control systems have been developed making use of customized Peltier assemblies, valve actuators, software, and amplification chemistry protocols. Multiple enhancements to the microfluidic chip design have been enacted to improve the reliability of sample delivery through the various on-chip modules. This work has been enabled by the encapsulation of PCR reagents into a solid phase material through an optimized Solid Phase Encapsulating Assay Mix (SPEAM) bead-based hydrogel fabrication process. SPEAM bead technology is reliably coupled with precise microfluidic metering and dispensing for efficient amplification and subsequent DNA short tandem repeat (STR) fragment analysis. This provides a means of on-chip reagent storage suitable for microfluidic automation, with the long shelf-life necessary for point-of-care (POC) or field deployable applications. This paper reports the first high quality 17-plex forensic STR amplification from a reference sample in a microfluidic chip with preloaded solid phase reagents, that is designed for integration with up and downstream processing.

show abstract

Human genomic DNA analysis using a semi‐automated sample preparation, amplification, and electrophoresis separation platform

Raisi¹,

Blizard²,

Shabari³

et al. 2004

J of Separation Science

Self Cite

View full text Add to dashboard Cite

The growing importance of analyzing the human genome to detect hereditary and infectious diseases associated with specific DNA sequences has motivated us to develop automated devices to integrate sample preparation, real-time PCR, and microchannel electrophoresis (MCE). In this report, we present results from an optimized compact system capable of processing a raw sample of blood, extracting the DNA, and performing a multiplexed PCR reaction. Finally, an innovative electrophoretic separation was performed on the post-PCR products using a unique MCE system. The sample preparation system extracted and lysed white blood cells (WBC) from whole blood, producing DNA of sufficient quantity and quality for a polymerase chain reaction (PCR). Separation of multiple amplicons was achieved in a microfabricated channel 30 microm x 100 microm in cross section and 85 mm in length filled with a replaceable methyl cellulose matrix operated under denaturing conditions at 50 degrees C. By incorporating fluorescent-labeled primers in the PCR, the amplicons were identified by a two-color (multiplexed) fluorescence detection system. Two base-pair resolution of single-stranded DNA (PCR products) was achieved. We believe that this integrated system provides a unique solution for DNA analysis.

show abstract

Toward a Rapid, Integrated, and Fully Automated DNA Diagnostic Assay for Chlamydia trachomatis and Neisseria gonorrhoeae

Cited by 13 publications

References 8 publications

Recent progress in the development of μTAS for clinical analysis

Recent progress in the development of μTAS for clinical analysis

Optimization of multiplexed PCR on an integrated microfluidic forensic platform for rapid DNA analysis

Human genomic DNA analysis using a semi‐automated sample preparation, amplification, and electrophoresis separation platform

Contact Info

Product

Resources

About