Acoustic detection of DNA conformation in genetic assays combined with PCR

Papadakis, George; Tsortos, Achilleas; Kordas, Antonis; Tiniakou, Ioanna; Morou, Evangelia; Vontas, John; Kardassis, Dimitris; Gizeli, Electra

doi:10.1038/srep02033

Cited by 30 publications

(32 citation statements)

References 35 publications

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“…There are reports about cDNA quantification and gene expression biosensor development. Han et al reported a cDNA quantification platform using a giant magnetoresistive (GMR) biosensor array [10], meanwhile Papadakis et al designed an electrochemical platform for direct detection of CYP707A1 gene expression from wheat leaves [11]. Davis et al developed a biosensor for SNP genotyping and gene expression quantification combining conventional PCR for target amplification and QCM for amplicon detection [12].…”

Section: Introductionmentioning

confidence: 99%

NLRP3, IL-1β, and Caspase-1 Gene Transcript Identification and Expression by QCM-D in Obese Children

et al. 2019

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Background. Obesity in children is highly prevalent in Mexican population. Adipose tissue has been related to specific pro- and anti-inflammatory cytokine and inflammasome gene and protein expression patterns. Actually, there is no existing biosensor for detecting gene expression patterns in children with obesity. The quartz crystal microbalance with dissipation monitoring (QCM-D) has been used as a transducer for DNA biosensor design. Results. In this study, the gene expression pattern of IL-1β, NLRP3, and CASPASE-1 in children with obesity was successfully determined by means of QCM-D. Gene expression patterns were validated with those obtained by quantitative polymerase chain reaction (qPCR), a validated molecular biology technique for gene expression quantification. QCM-D analysis of the detected mass corresponding results for each of the genes showed a major detected mass for IL-1β, followed by similar NLRP3 and constitutive gene 18S deposited mass and a smaller deposited mass for CASPASE-1. Surprisingly, when comparing mRNA gene expression results for NLRP3, IL-1β, and CASPASE-1 obtained with qPCR and QCM-D, similar patterns were found, revealing greatest expression of IL-1β, followed by NLRP3, with CASPASE-1 being the molecule of least expression in the group of children with obesity. AFM images illustrate the step-by-step changes that took place on the quartz surface. Conclusions. QCM-D proved successfully for determining the gene transcripts and expression of NLRP3, IL-1β, and CASPASE-1 in children with obesity, with similar results validated by qPCR. “QCM-D decreases detection costs compared with a validated molecular biology technique.” The QCM-D biosensor developed by our group was successful for gene expression determination; in the future, it can be used for molecular diagnosis.

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Section: Introductionmentioning

confidence: 99%

NLRP3, IL-1β, and Caspase-1 Gene Transcript Identification and Expression by QCM-D in Obese Children

et al. 2019

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“…Throat swab specimens were obtained on day 6 of illness, and rectal swab specimens were obtained on day 8 of illness. Specimens were tested for enterovirus and HPeV RNA by reverse transcription PCR (RT-PCR) and primers specific for the highly conserved 5′ untranslated region ( 1 ) (details for HPeV primers and probes are available on request). HPeV was detected in the throat swab specimen, but not the rectal swab specimen.…”

Section: The Studymentioning

confidence: 99%

“…We attempted molecular typing of HPeV by using the method of Papadakis et al ( 1 ) and primers AN353, AN355, AN357, AN358, and AN369 described by Nix et al ( 2 ). However, typing was not successful because of low copy numbers, probably caused by specimens being collected late in the illness.…”

Section: The Studymentioning

confidence: 99%

Myocarditis Caused by Human Parechovirus in Adult

Kong¹,

Lau²,

Goh³

et al. 2017

Emerg. Infect. Dis.

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“…In a recent study, the polymerase chain reaction was paired with acoustic biosensors for detecting different types of mutations in Anopheles and the human BRCA1 and BRCA2 genes [ 7 , 8 ]. The described methodology combines the sensitivity and selectivity of the PCR method with the label free nature of acoustic biosensing while DNA detection is achieved within only a few minutes and during real time monitoring.…”

Section: Introductionmentioning

confidence: 99%

Bacteria Murmur: Application of an Acoustic Biosensor for Plant Pathogen Detection

et al. 2015

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A multi-targeting protocol for the detection of three of the most important bacterial phytopathogens, based on their scientific and economic importance, was developed using an acoustic biosensor (the Quartz Crystal Microbalance) for DNA detection. Acoustic detection was based on a novel approach where DNA amplicons were monitored and discriminated based on their length rather than mass. Experiments were performed during real time monitoring of analyte binding and in a direct manner, i.e. without the use of labels for enhancing signal transduction. The proposed protocol improves time processing by circumventing gel electrophoresis and can be incorporated as a routine detection method in a diagnostic lab or an automated lab-on-a-chip system for plant pathogen diagnostics.

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Acoustic detection of DNA conformation in genetic assays combined with PCR

Cited by 30 publications

References 35 publications

NLRP3, IL-1β, and Caspase-1 Gene Transcript Identification and Expression by QCM-D in Obese Children

NLRP3, IL-1β, and Caspase-1 Gene Transcript Identification and Expression by QCM-D in Obese Children

Myocarditis Caused by Human Parechovirus in Adult

Bacteria Murmur: Application of an Acoustic Biosensor for Plant Pathogen Detection

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