Miniature on-chip detection of unpurified methicillin-resistant Staphylococcus aureus (MRSA) DNA using real-time PCR

House, Dustin; Chon, Chan Hee; Creech, C. Buddy; Skaar, Eric P.; Li, Dongqing

doi:10.1016/j.jbiotec.2009.12.013

Cited by 26 publications

(10 citation statements)

References 32 publications

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“…The development of labon-a-chip (LOC) devices, a term for "micro-total analysis systems (µTAS)", for biochemical analyses has seen explosive growth over the past decade. [6][7][8] LOC devices are now widely used in biology and biotechnology, 9 DNA and proteins analysis, 10,11 high throughput screening 12 and chemical reactions. 13 Various bio-substances are often analyzed by detecting the immune reaction between antigens in a sample solution or antibodies immobilized on micro channel surfaces using fluorescence detection and the surface Plasmon resonance method.…”

Section: Introductionmentioning

confidence: 99%

Simple Potential Bioanalysis Micro reactor Fabricated By Vertically-Aligned Multi-Walled Carbon Nanotubes (MWCNTs)

Guo

Tam

2017

JABB

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Section: Introductionmentioning

confidence: 99%

Simple Potential Bioanalysis Micro reactor Fabricated By Vertically-Aligned Multi-Walled Carbon Nanotubes (MWCNTs)

Guo

Tam

2017

JABB

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“…21 The approach of separating disposable chip and reusable heating units is often seen in plastic PCR chips. [22][23][24][25][26] One reason for this is that conventional MEMS manufacturing technologies are less compatible with common lab-on-chip substrates making integration of electronic features sometimes difficult in polymer systems. Using external heating, on the one hand reduces the number of fabrication steps of the chip, but on the other hand, makes thermal contact between the chip and heating block, and heat transfer through plastic materials with low thermal conductance, a critical issue to obtain fast heating rates and thermal accuracy.…”

Section: Introductionmentioning

confidence: 99%

Genotyping of single nucleotide polymorphisms by melting curve analysis using thin film semi-transparent heaters integrated in a lab-on-foil system

et al. 2013

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The recent technological advances in micro/nanotechnology present new opportunities to combine microfluidics with microarray technology for the development of small, sensitive, single-use, point-of-care molecular diagnostic devices. As such, the integration of microarray and plastic microfluidic systems is an attractive low-cost alternative to glass based microarray systems. This paper presents the integration of a DNA microarray and an all-polymer microfluidic foil system with integrated thin film heaters, which demonstrate DNA analysis based on melting curve analysis (MCA). A novel micro-heater concept using semi-transparent copper heaters manufactured by roll-to-roll and lift-off on polyethylene naphthalate (PEN) foil has been developed. Using a mesh structure, heater surfaces have been realized in only one single metallization step, providing more efficient and homogenous heating characteristics than conventional meander heaters. A robust DNA microarray spotting protocol was adapted on Parylene C coated heater-foils, using co-polymer poly(DMA-NAS-MAPS) to enable covalent immobilization of DNA. The heaters were integrated in a microfluidic channel using lamination foils and MCA of the spotted DNA duplexes showed single based discrimination of mismatched over matched target DNA-probes. Finally, as a proof of principle, we perform MCA on PCR products to detect the Leu7Pro polymorphism of the neutropeptide Y related to increased risk of Type II diabetes, BMI and depression.

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“…Ethidium monoazide (EMA) is a cell membrane impermeable, fluorescent, nucleic acid staining dye. 22 The intermediate compound, nitrene, was produced by photolysis of EMA with visible light exposure that could be bound to DNA to form a stable monoadduct. 23 Free EMA in solution was photolyzed and converted to hydroxylamine that lost covalent bonding capability with DNA.…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of live methicillin-resistant Staphylococcus aureus by using an integrated microfluidic system capable of ethidium monoazide pre-treatment and molecular diagnosis

Liu

Wang

et al. 2012

Biomicrofluidics

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Methicillin-resistant Staphylococcus aureus (MRSA) is a bacterium resistant to all existing penicillin and lactam-based antimicrobial drugs and, therefore, has become one of the most prevalent antibiotic-resistant pathogens found in hospitals. The multi-drug resistant characteristics of MRSA make it challenging to clinically treat infected patients. Therefore, early diagnosis of MRSA has become a public-health priority worldwide. Conventionally, cell-culture based methodology and microscopic identification are commonly used for MRSA detection. However, they are relatively time-consuming and labor-intensive. Recently, molecular diagnosis based on nucleic acid amplification techniques, such as polymerase chain reaction (PCR), has been widely investigated for the rapid detection of MRSA. However, genomic DNA of both live and dead pathogens can be distinguished by conventional PCR. These results thus could not provide sufficient confirmation of an active infection for clinicians. In this study, live MRSA was rapidly detected by using a new integrated microfluidic system. The microfluidic system has been demonstrated to have 100% specificity to detect live MRSA with S. aureus and other pathogens commonly found in hospitals. The experimental results showed that the limit of detection for live MRSA from biosamples was approximately 10(2) CFU/μl. In addition, the entire diagnostic protocol, from sample pre-treatment to fluorescence observation, can be automatically completed within 2.5 h. Consequently, this microfluidic system may be a powerful tool for the rapid molecular diagnosis of live MRSA.

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Miniature on-chip detection of unpurified methicillin-resistant Staphylococcus aureus (MRSA) DNA using real-time PCR

Cited by 26 publications

References 32 publications

Simple Potential Bioanalysis Micro reactor Fabricated By Vertically-Aligned Multi-Walled Carbon Nanotubes (MWCNTs)

Simple Potential Bioanalysis Micro reactor Fabricated By Vertically-Aligned Multi-Walled Carbon Nanotubes (MWCNTs)

Genotyping of single nucleotide polymorphisms by melting curve analysis using thin film semi-transparent heaters integrated in a lab-on-foil system

Rapid detection of live methicillin-resistant Staphylococcus aureus by using an integrated microfluidic system capable of ethidium monoazide pre-treatment and molecular diagnosis

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