Sensitive Molecular Diagnostics Using Volume-Amplified Magnetic Nanobeads

Strömberg, Mattias; Göransson, Jenny; Gunnarsson, Klas; Nilsson, Mats; Svedlindh, Peter; Strömme, Maria

doi:10.1021/nl072760e

Cited by 118 publications

(153 citation statements)

References 25 publications

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“…Padlock probes are an alternative to PCR. They can be highly multiplex and can be combined with magnetic nanoparticles (17) in a commercial portable read-out device (24), although the short shanks make them especially vulnerable to pathogen variation. There is room for a new principle for the multiplex detection of nucleic acids.…”

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confidence: 99%

Variation-Tolerant Capture and Multiplex Detection of Nucleic Acids: Application to Detection of Microbes

et al. 2012

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In contrast to ordinary PCRs, which have a limited multiplex capacity and often return false-negative results due to target variation or inhibition, our new detection strategy, VOCMA (variation-tolerant capture multiplex assay), allows variation-tolerant, target-specific capture and detection of many nucleic acids in one test. Here we demonstrate the use of a single-tube, dual-step amplification strategy that overcomes the usual limitations of PCR multiplexing, allowing at least a 22-plex format with retained sensitivity. Variation tolerance was achieved using long primers and probes designed to withstand variation at known sites and a judicious mix of degeneration and universal bases. We tested VOCMA in situations where enrichment from a large sample volume with high sensitivity and multiplexity is important (sepsis; streptococci, enterococci, and staphylococci, several enterobacteria, candida, and the most important antibiotic resistance genes) and where variation tolerance and high multiplexity is important (gastroenteritis; astrovirus, adenovirus, rotavirus, norovirus genogroups I and II, and sapovirus, as well as enteroviruses, which are not associated with gastroenteritis). Detection sensitivities of 10 to 1,000 copies per reaction were achieved for many targets. VOCMA is a highly multiplex, variation-tolerant, general purpose nucleic acid detection concept. It is a specific and sensitive method for simultaneous detection of nucleic acids from viruses, bacteria, fungi, and protozoa, as well as host nucleic acid, in the same test. It can be run on an ordinary PCR and a Luminex machine and is suitable for both clinical diagnoses and microbial surveillance.

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Variation-Tolerant Capture and Multiplex Detection of Nucleic Acids: Application to Detection of Microbes

et al. 2012

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“…In previous studies that involve C2CA technology, RCA products were detected by radiation-labeled detection oligonucleotides, fluorescence-labeled detection oligonucleotides, or paramagnetic beads (4,11,19). However, these methods often require advanced and expensive instrumentation, which made them unsuitable for field-based diagnosis.…”

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Colorimetric Nucleic Acid Testing Assay for RNA Virus Detection Based on Circle-to-Circle Amplification of Padlock Probes

Zorzet

Göransson

et al. 2011

J Clin Microbiol

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We developed a molecular diagnostic method for detection of RNA virus based on padlock probes and colorimetric readout. The feasibility of our approach was demonstrated by using detection of Crimean-Congo hemorrhagic fever (CCHF) virus as a model. Compared with conventional PCR-based methods, our approach does not require advanced equipment, involves easier assay design, and has a sensitivity of 10 3 viral copies/ml. By using a cocktail of padlock probes, synthetic templates representing different viral strain variants could be detected. We analyzed 34 CCHF patient samples, and all patients were correctly diagnosed when the results were compared to those of the current real-time PCR method. This is the first time that highly specific padlock probes have been applied to detection of a highly variable target sequence typical of RNA viruses.

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“…This method uses the fact that the increase of the hydrodynamic volume of a magnetic particle bound to a biological target results in a shift of the Brownian relaxation frequency relative to that for particles not bound to the biological target. This shift can be large enough to be registered by a magnetic field sensor [27,28], Fig. 1.…”

Section: Fig 1 Dynamic Field Range and Detectivity Of Some Magnetommentioning

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Modelling and design of planar Hall effect bridge sensors for low-frequency applications

Persson

Bejhed

Østerberg

et al. 2013

Sensors and Actuators A: Physical

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This is an accepted version of a paper published in Sensors and Actuators A-Physical. This paper has been peer-reviewed but does not include the final publisher proofcorrections or journal pagination.Citation for the published paper: Persson, A., Bejhed, R., Oesterberg, F., Gunnarsson, K., Nguyen, H. et al. (2012) "Modelling and design of planar Hall effect bridge sensors for low-frequency applications" Sensors and Actuators Access to the published version may require subscription. AbstractThe applicability of miniaturized magnetic field sensors is being explored in several areas of magnetic field detection due to their integratability, low mass, and potentially low cost. In this respect, different thin-film technologies, especially those employing magnetoresistance, show great potential, being compatible with batch micro-and nanofabrication techniques. For lowfrequency magnetic field detection, sensors based on the planar Hall effect, especially planar Hall effect bridge (PHEB) sensors, show promising performance given their inherent lowfield linearity, limited hysteresis and moderate noise figure. In this work, the applicability of such PHEB sensors to different areas is investigated. An analytical model is constructed to estimate the performance of an arbitrary PHEB sensor geometry in terms of, e.g., sensitivity and detectivity. The model is valid for an ideal case, e.g., disregarding shape anisotropy effects, and also incorporates some approximations. To validate the results, modelled data was compared to measurements on actual PHEBs and was found to predict the measured values within 13% for the investigated geometries. Subsequently, the model was used to establish a design process for optimizing a PHEB to a particular set of requirements on the bandwidth, detectivity, compliance voltage and amplified signal-to-noise ratio. By applying this design process, the size, sensitivity, resistance, bias current and power consumption of the PHEB can be estimated. The model indicates that PHEBs can be applicable to several different areas within science including satellite attitude determination and magnetic bead detection in labon-a-chip applications, where detectivities down towards 1 nTHz -0.5 at 1 Hz are required, and maybe even magnetic field measurements in scientific space missions and archaeological surveying, where the detectivity has to be less than 100 pTHz -0.5 at 1 Hz.

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Sensitive Molecular Diagnostics Using Volume-Amplified Magnetic Nanobeads

Cited by 118 publications

References 25 publications

Variation-Tolerant Capture and Multiplex Detection of Nucleic Acids: Application to Detection of Microbes

Variation-Tolerant Capture and Multiplex Detection of Nucleic Acids: Application to Detection of Microbes

Colorimetric Nucleic Acid Testing Assay for RNA Virus Detection Based on Circle-to-Circle Amplification of Padlock Probes

Modelling and design of planar Hall effect bridge sensors for low-frequency applications

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