Taking It to the Extreme: PCR at Wittwerspeed

Mackay, John F.

doi:10.1373/clinchem.2014.233338

Cited by 6 publications

(5 citation statements)

References 13 publications

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“…It is both reliable and reproducible but also time‐consuming and poorly adapted to clinical application, especially when a rapid result is required. Automated RNA isolation and RT solutions, as well as ultra‐fast PCR, could improve RT‐qPCR applicability . Moreover, novel probe‐based technologies are emerging and may allow quantitative measures that are much faster and easier to use in a clinical context .…”

Section: Circulating Myomirs In Cancermentioning

confidence: 99%

“…Automated RNA isolation and RT solutions, as well as ultrafast PCR, could improve RT-qPCR applicability. [62][63][64] Moreover, novel probe-based technologies are emerging and may allow quantitative measures that are much faster and easier to use in a clinical context. [65][66][67] Third, we need to determine normal values in a wide population of healthy individuals in order to identify inter-individual variability with regards to gender, age, ethnicity, or training status.…”

Section: Considerations For Clinical Applicationmentioning

confidence: 99%

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Circulating myomiRs: a new class of biomarkers to monitor skeletal muscle in physiology and medicine

Siracusa¹,

Koulmann

Banzet

2017

J cachexia sarcopenia muscle

107

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MicroRNAs (miRNA) are small non‐coding RNAs that target mRNAs and are consequently involved in the post‐transcriptional regulation of gene expression. Some miRNAs are ubiquitously expressed in tissue, while others are tissue‐specific or tissue‐enriched. miRNAs can be released by cells and are found in various biofluids, including serum and plasma. Thus, measuring miRNAs in the circulation may provide information on the originating tissue or cells. MyomiRs are described as striated muscle‐specific or muscle‐enriched miRNAs. Their circulating levels can be measured and have been proposed to be new biomarkers of physiological and pathological muscle processes. The aims of this review are to summarize the current knowledge of circulating myomiRs, to identify the types of information they can provide about skeletal muscle, and to determine how to apply that information in the fields of research and medicine.

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Section: Circulating Myomirs In Cancermentioning

confidence: 99%

Section: Considerations For Clinical Applicationmentioning

confidence: 99%

Circulating myomiRs: a new class of biomarkers to monitor skeletal muscle in physiology and medicine

Siracusa¹,

Koulmann

Banzet

2017

J cachexia sarcopenia muscle

107

View full text Add to dashboard Cite

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“…He accomplishes the required speeds by using 2 large water baths to rapidly change the temperature. As noted in this journal, this design is "harking back to the early days of PCR" (6 ).…”

Section: How Does It Work?mentioning

confidence: 99%

Extreme PCR: A Breakthrough Innovation for Outbreaks?

Kumar

Webster²

2015

Clinical Chemistry

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“…With techniques available that perform nucleic acid extraction in less than 10 min, ultra-fast (in under 5 min) quantitative PCR (UF-qPCR) is a target to effectively allow while-you-wait testing [4][5][6]. As noted previously [7], the present challenge for decreasing PCR amplification times lies with instrumentation, not with PCR chemistry.…”

Section: Introductionmentioning

confidence: 99%

Ultra-fast, sensitive and quantitative on-chip detection of group B streptococci in clinical samples

Cai

Fauvart

Wiederkehr

et al. 2019

Talanta

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PCR enables sensitive and specific detection of infectious disease agents, but application in pointof-care diagnostic testing remains scarce. A compact tool that runs PCR assays in less than a few minutes and that relies on mass-producible, disposable reactors could revolutionize while-you-wait molecular testing. We here exploit well-established semiconductor manufacturing processes to produce silicon ultra-fast quantitative PCR (UF-qPCR) chips that can run PCR protocols with limited assay optimization. A total of 110 clinical samples were analyzed for the detection of group B streptococci using both a validated benchtop and an on-chip qPCR assay. For the on-chip assay, the total reaction time was reduced after optimization to less than 5 min. The standard curve, spanning a concentration range of 5 log units, yielded a PCR efficiency of 94%. The sensitivity obtained was 96% (96/100; CI: 90-98%) and the specificity 70% (7/10; CI: 40-90%). As shown, in case melting analyses would be integrated, the obtained sensitivity would slightly drop to 93% (CI: 86%-96%), while the specificity would increase to 100% (CI: 72% -100%). In comparison to the benchtop reference qPCR assay performed on a LightCycler © 96, the on-chip assay demonstrated a highly significant qualitative (Spearman's rank correlation) and quantitative (linear regression) correlation. Using a mass-producible qPCR chip and limited assay optimization, we were able to develop a validated qPCR protocol that can be carried out in less than five minutes. The analytical performance of the microchip-based UF-qPCR system was shown to match that of a benchtop assay. This is the first report to provide UF-qPCR validation using clinical samples. We demonstrate that qPCR-based while-you-wait testing is feasible without jeopardizing assay performance.

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Taking It to the Extreme: PCR at Wittwerspeed

Cited by 6 publications

References 13 publications

Circulating myomiRs: a new class of biomarkers to monitor skeletal muscle in physiology and medicine

Circulating myomiRs: a new class of biomarkers to monitor skeletal muscle in physiology and medicine

Extreme PCR: A Breakthrough Innovation for Outbreaks?

Ultra-fast, sensitive and quantitative on-chip detection of group B streptococci in clinical samples

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