Bacterial RNA Extraction and Purification from Whole Human Blood Using Isotachophoresis

Rogacs, Anita; Qu, Yatian; Santiago, Juan G.

doi:10.1021/ac301021d

Cited by 43 publications

(42 citation statements)

References 56 publications

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“…A modified CE-based method, isotachophoresis (ITP), is a simpler design (Figure 4(b)) relying on similar CE separation strategies (Persat et al 2009; Young et al 2010; Jung et al 2011; Garcia-Schwarz et al 2012). This versatile platform has been developed into a LOC, and has a wide range of nucleic acid separation for diagnostic applications such as RNA extraction from bacteria (Rogacs et al 2012), DNA isolation for the Hepatitis B virus (Liu et al 2006) and DNA extraction of the Malaria parasite, Plasmodium from infected erythrocytes (Marshall et al 2011). Once isolated, the nucleic acid can also be amplified on-a-chip (Liu et al 2006) using DNA amplification methods.…”

Section: Development Of Potential Alternative and Future Biosensors Umentioning

confidence: 99%

Strategies in Ebola virus disease (EVD) diagnostics at the point of care

Coarsey

Esiobu

Narayanan

et al. 2017

Critical Reviews in Microbiology

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Ebola virus disease (EVD) is a devastating, highly infectious illness with a high mortality rate. The disease is endemic to regions of Central and West Africa, where there is limited laboratory infrastructure and trained staff. The recent 2014 West African EVD outbreak has been unprecedented in case numbers and fatalities, and has proven that such regional outbreaks can become a potential threat to global public health, as it became the source for the subsequent transmission events in Spain and the USA. The urgent need for rapid and affordable means of detecting Ebola is crucial to control the spread of EVD and prevent devastating fatalities. Current diagnostic techniques include molecular diagnostics and other serological and antigen detection assays; which can be time-consuming, laboratory-based, often require trained personnel and specialized equipment. In this review, we discuss the various Ebola detection techniques currently in use, and highlight the potential future directions pertinent to the development and adoption of novel point-of-care diagnostic tools. Finally, a case is made for the need to develop novel microfluidic technologies and versatile rapid detection platforms for early detection of EVD.

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Section: Development Of Potential Alternative and Future Biosensors Umentioning

confidence: 99%

Strategies in Ebola virus disease (EVD) diagnostics at the point of care

Coarsey

Esiobu

Narayanan

et al. 2017

Critical Reviews in Microbiology

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“…ITP uses a heterogeneous buffer system consisting of high-mobility ions in the leading electrolyte and low-mobility ions in the terminating electrolyte; the analytes of interest have intermediate ionic mobility. An applied electric potential separates the ionic species based on their electrophoretic mobility, thus focusing the analytes at the leading and terminating electrolyte interface (Figure 6) (56).The efficacy and versatility of this technique has been demonstrated in the purification of genomic DNA from WB (57), the isolation of small RNA from cell lysate (58), the extraction of DNA from malaria-infected erythrocytes (59), as well as the purification of bacterial RNA from WB lysate (60). Additionally, nucleic acid detection has been implemented with ITP using loop-mediated isothermal amplification (a process known as NAIL) on an integrated chip to detect pathogenic Escherichia coli O157:H7 cells from whole milk (61).…”

Section: Isotachophoresismentioning

confidence: 99%

Microfluidic Sample Preparation for Medical Diagnostics

Cui

Rhee

Singh

et al. 2015

Annu. Rev. Biomed. Eng.

111

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Fast and reliable diagnoses are invaluable in clinical care. Samples (e.g., blood, urine, and saliva) are collected and analyzed for various biomarkers to quickly and sensitively assess disease progression, monitor response to treatment, and determine a patient's prognosis. Processing conventional samples entails many manual time-consuming steps. Consequently, clinical specimens must be processed by skilled technicians before antigens or nucleic acids are detected, and these are often present at dilute concentrations. Recently, several automated microchip technologies have been developed that potentially offer many advantages over traditional bench-top extraction methods. The smaller length scales and more refined transport mechanisms that characterize these microfluidic devices enable faster and more efficient biomarker enrichment and extraction. Additionally, they can be designed to perform multiple tests or experimental steps on one integrated, automated platform. This review explores the current research on microfluidic methods of sample preparation that are designed to aid diagnosis, and covers a broad spectrum of extraction techniques and designs for various types of samples and analytes.

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“…Santiago's group has demonstrated that a sample can be continuously injected from the TE, rather than changing to a separate TE vial [28][29][30][31][32]. When compared to a discrete fixed-time injection of the sample, this should help to provide lower detection limits.…”

Section: Continuous Injectionmentioning

confidence: 99%

Counter-pressure-assisted ITP with electrokinetic injection under field-amplified conditions for bacterial analysis

et al. 2015

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Counter-pressure was used to extend the duration of field-amplified sample injection in isotachophoresis (FASI-ITP) in order to improve the detection of bacterial cells. Using 0.51-μm negatively charged encapsulated fluorescent beads as a model, the counter-pressure, injection and separation voltages, and times were optimized. Using 6-min 8,963-Pa counter-pressure FASI-ITP injections at -12 kV followed by mobilization of the ITP band with continued injection at -6 kV, the limit of detection (LOD) for Escherichia coli was improved to 78 cells/mL, a factor of 4 when compared with FASI-ITP without counter-pressure.

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Bacterial RNA Extraction and Purification from Whole Human Blood Using Isotachophoresis

Cited by 43 publications

References 56 publications

Strategies in Ebola virus disease (EVD) diagnostics at the point of care

Strategies in Ebola virus disease (EVD) diagnostics at the point of care

Microfluidic Sample Preparation for Medical Diagnostics

Counter-pressure-assisted ITP with electrokinetic injection under field-amplified conditions for bacterial analysis

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