Multiplexed Point-of-Care Testing – xPOCT

Dincer, Can; Bruch, Richard C.; Kling, André; Dittrich, Petra S.; Urban, G.

doi:10.1016/j.tibtech.2017.03.013

Cited by 428 publications

(292 citation statements)

References 84 publications

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“…The authors further claimed that partial automation of the real-time PCR assay allowed testing of approximately 200 samples per day. However, the trend in infectious disease diagnosis is toward full automation, multiplexing, and miniaturization of assay systems to facilitate rapid point-of-care (POC) diagnosis (8,9). It is encouraging to note that another group has successfully FIG 1 BD Max Candida auris assay sensitivity.…”

Section: Discussionmentioning

confidence: 99%

A Rapid and Automated Sample-to-Result Candida auris Real-Time PCR Assay for High-Throughput Testing of Surveillance Samples with the BD Max Open System

et al. 2019

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The multidrug-resistant yeast pathogen Candida auris continues to cause outbreaks and clusters of clinical cases worldwide. Previously, we developed a real-time PCR assay for the detection of C. auris from surveillance samples (L. Leach, Y. Zhu, and S. Chaturvedi, J Clin Microbiol 56:e01223-17, 2018, https://doi.org/10.1128/ JCM.01223-17). The assay played a crucial role in the ongoing investigations of the C. auris outbreak in New York City. To ease the implementation of the assay in other laboratories, we developed an automated sample-to-result real-time C. auris PCR assay using the BD Max open system. We optimized sample extraction at three different temperatures and four incubation periods. Sensitivity was determined using eight pools of patient samples, and specificity was calculated using four clades of C. auris and closely and distantly related yeasts. Three independent extractions and testing of two patient sample pools in quadruplicate yielded assay precision. BD Max optimum assay conditions were as follows: DNA extraction at 75°C for 20 min and the use of PerfeCTa multiplex quantitative PCR (qPCR) ToughMix. The limit of detection (LOD) of the assay was one C. auris CFU/PCR. We detected all four clades of C. auris without cross-reactivity to other yeasts. Of the 110 patient surveillance samples tested, 50 were positive for C. auris using the BD Max system with 96% clinical sensitivity and 94% accuracy compared to the results of the manual assay. The BD Max assay allows high-throughput C. auris screening of 180 surveillance samples in a 12-h workday. C andida auris, an emerging multidrug-resistant yeast pathogen, continues to cause outbreaks and clusters of clinical cases worldwide (1, 2). There are ongoing efforts to devise better diagnostic approaches for the rapid detection of C. auris in clinical and surveillance samples (3-5). Previously, we developed and validated a manual real-time PCR assay for the direct detection of C. auris from surveillance samples at the New York State Department of Health (NYSDOH) Mycology Laboratory (3). The laboratorydeveloped test (LDT) enabled NYSDOH laboratory scientists and epidemiologists to carry out unprecedented surveillance and testing in hospitals and health care facilities in New York City. To date, over 13,000 clinical samples from 151 health care facilities and over 1,000 C. auris isolates were processed (S. Chaturvedi, unpublished data). The Candida auris LDT, standard operating procedures, and validation results were shared extensively with hospital, commercial, and public health laboratories. However, the C. auris LDT is not amenable to automation and high-throughput screening, and adoption of the LDT has progressed slowly while the affected facilities and sample numbers continue to grow. Therefore, we developed and validated a real-time PCR assay for C. Citation Leach L, Russell A, Zhu Y, Chaturvedi S, Chaturvedi V. 2019. A rapid and automated sample-to-result Candida auris real-time PCR assay for high-throughput testing of surveillance samples with the B...

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Section: Discussionmentioning

confidence: 99%

A Rapid and Automated Sample-to-Result Candida auris Real-Time PCR Assay for High-Throughput Testing of Surveillance Samples with the BD Max Open System

et al. 2019

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“…However, these approaches still need error‐prone amplification steps prior to the miRNA detection, have rather tedious preparation steps, or are limited by the design of suitable primers . To address these issues, we combine microfluidics with an electrochemical signal readout to develop a sensitive (i.e., target amplification‐free) and selective diagnostic test, while enabling a miniaturization for POC testing . We further apply the newly discovered CRISPR/Cas13a technology, which is able of targeting almost any RNA, to our developed electrochemical biosensor, creating a novel and powerful tool for miRNA diagnostics ( Figure a).…”

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confidence: 99%

CRISPR/Cas13a‐Powered Electrochemical Microfluidic Biosensor for Nucleic Acid Amplification‐Free miRNA Diagnostics

et al. 2019

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Noncoding small RNAs, such as microRNAs, are becoming the biomarkers of choice for multiple diseases in clinical diagnostics. A dysregulation of these microRNAs can be associated with many different diseases, such as cancer, dementia, and cardiovascular conditions. The key for effective treatment is an accurate initial diagnosis at an early stage, improving the patient's survival chances. In this work, the first clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a‐powered microfluidic, integrated electrochemical biosensor for the on‐site detection of microRNAs is introduced. Through this unique combination, the quantification of the potential tumor markers microRNA miR‐19b and miR‐20a is realized without any nucleic acid amplification. With a readout time of 9 min and an overall process time of less than 4 h, a limit of detection of 10 pm is achieved, using a measuring volume of less than 0.6 µL. Furthermore, the feasibility of the biosensor platform to detect miR‐19b in serum samples of children, suffering from brain cancer, is demonstrated. The validation of the obtained results with a standard quantitative real‐time polymerase chain reaction method shows the ability of the electrochemical CRISPR‐powered system to be a low‐cost, easily scalable, and target amplification‐free tool for nucleic acid based diagnostics.

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“…371 There is an urgent need to continue progress towards developing platforms capable of multiplexing disease targets into a single test without sacrificing test sensitivity. 372 …”

Section: Discussionmentioning

confidence: 99%

Point-of-care diagnostics to improve maternal and neonatal health in low-resource settings

et al. 2017

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Each day, approximately 830 women and 7,400 newborns die from complications during pregnancy and childbirth. Improving maternal and neonatal health will require bringing rapid diagnosis and treatment to the point of care in low-resource settings. However, to date there are few diagnostic tools available that can be used at the point of care to detect the leading causes of maternal and neonatal mortality in low-resource settings. Here we review both commercially available diagnostics and technologies that are currently in development to detect the leading causes of maternal and neonatal mortality, highlighting key gaps in development where innovative design could increase access to technology and enable rapid diagnosis at the bedside.

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Multiplexed Point-of-Care Testing – xPOCT

Cited by 428 publications

References 84 publications

A Rapid and Automated Sample-to-Result Candida auris Real-Time PCR Assay for High-Throughput Testing of Surveillance Samples with the BD Max Open System

A Rapid and Automated Sample-to-Result Candida auris Real-Time PCR Assay for High-Throughput Testing of Surveillance Samples with the BD Max Open System

CRISPR/Cas13a‐Powered Electrochemical Microfluidic Biosensor for Nucleic Acid Amplification‐Free miRNA Diagnostics

Point-of-care diagnostics to improve maternal and neonatal health in low-resource settings

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