A compact, low-cost, quantitative and multiplexed fluorescence detection platform for point-of-care applications

Smith, John E.; Zhu, Meilin; Katchman, Benjamin A.; Arafa, Hany; Anderson, Karen S.; Christen, Jennifer Blain

doi:10.1016/j.bios.2018.04.002

Cited by 43 publications

(17 citation statements)

References 20 publications

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“…Even though they report results within 5 to 15 min, they commonly suffer from low sensitivity or low specificity for clinical applications. In addition, they mainly rely on colorimetric or fluorescence measurements which are not usually capable of quantification unless expensive equipment such as optical cameras or lasers are involved [11]. Instead, nucleic-acid amplification tests (NAATs) are characterized by high sensitivity and specificity, enabling quantification and detection of early stage infections.…”

Section: Introductionmentioning

confidence: 99%

Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform

Malpartida-Cardenas

Miscourides

Rodríguez-Manzano

et al. 2019

Biosensors and Bioelectronics

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Early and accurate diagnosis of malaria and drug-resistance is essential to effective disease management. Available rapid malaria diagnostic tests present limitations in analytical sensitivity, drug-resistance testing and/or quantification. Conversely, diagnostic methods based on nucleic acid amplification stepped forwards owing to their high sensitivity, specificity and robustness. Nevertheless, these methods commonly rely on optical measurements and complex instrumentation which limit their applicability in resource-poor, point-of-care settings. This paper reports the specific, quantitative and fully-electronic detection of Plasmodium falciparum, the predominant malaria-causing parasite worldwide, using a Lab-on-Chip platform developed inhouse. Furthermore, we demonstrate on-chip detection of C580Y, the most prevalent singlenucleotide polymorphism associated to artemisinin-resistant malaria. Real-time non-optical DNA sensing is facilitated using Ion-Sensitive Field-Effect Transistors, fabricated in unmodified complementary metal-oxide-semiconductor (CMOS) technology, coupled with loop-mediated isothermal amplification. This work holds significant potential for the development of a fully portable and quantitative malaria diagnostic that can be used as a rapid point-of-care test.

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Section: Introductionmentioning

confidence: 99%

Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform

Malpartida-Cardenas

Miscourides

Rodríguez-Manzano

et al. 2019

Biosensors and Bioelectronics

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“…These foils are comparably thin, allowing to reduce the optical path to about 2 mm in total. Conceptually related approaches have been described for using polarizers [9] and low-cost bandpass filters [11], respectively.…”

Section: Resultsmentioning

confidence: 99%

A low-cost fluorescence reader for in vitro transcription and nucleic acid detection with Cas13a

et al. 2019

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Point-of-care testing (POCT) in low-resource settings requires tools that can operate independently of typical laboratory infrastructure. Due to its favorable signal-to-background ratio, a wide variety of biomedical tests utilize fluorescence as a readout. However, fluorescence techniques often require expensive or complex instrumentation and can be difficult to adapt for POCT. To address this issue, we developed a pocket-sized fluorescence detector costing less than $15 that is easy to manufacture and can operate in low-resource settings. It is built from standard electronic components, including an LED and a light dependent resistor, filter foils and 3D printed parts, and reliably reaches a lower limit of detection (LOD) of ≈ 6.8 nM fluorescein, which is sufficient to follow typical biochemical reactions used in POCT applications. All assays are conducted on filter paper, which allows for a flat detector architecture to improve signal collection. We validate the device by quantifying in vitro RNA transcription and also demonstrate sequence-specific detection of target RNAs with an LOD of 3.7 nM using a Cas13a-based fluorescence assay. Cas13a is an RNA-guided, RNA-targeting CRISPR effector with promiscuous RNase activity upon recognition of its RNA target. Cas13a sensing is highly specific and adaptable and in combination with our detector represents a promising approach for nucleic acid POCT. Furthermore, our open-source device may be used in educational settings, through providing low cost instrumentation for quantitative assays or as a platform to integrate hardware, software and biochemistry concepts in the future.

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“…It can be used to determine the efficiency of detection. Therefore, colored microspheres for tracers based on immunochromatographic assays have gradually attracted attention in the medical science and food security fields [24–26]. In addition, real-time RT-PCR was considered as a reference method used to detect the different results of FMT and CTM.…”

Section: Discussionmentioning

confidence: 99%

Comparison of the detection performance of two different one-step-combined test strips with fluorescent microspheres or colored microspheres as tracers for influenza A and B viruses

Liao

Wang

et al. 2019

Virol J

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Background Influenza A and B viruses mainly cause respiratory infectious disease. Till now, few tests are able to simultaneously detect both, especially in primary medical establishments. Methods This study was designed to compare the performance of two different one-step-combined test strips for the detection of influenza A and B: one strip with fluorescent microspheres for tracers (FMT); and the other strip with colored microspheres for tracers (CMT). To test the strips, cultures of influenza A, B, and other pathogenic viruses were used, in addition to 1085 clinical specimens from symptomatic patients with respiratory infections. Real-time RT-PCR was also considered as a reference method used to detect the different results of FMT and CTM. Results Detection thresholds for influenza A and B cultures using serial dilutions revealed that the sensitivity of FMT was higher than that of CMT (both P < 0.05). With the culture mixtures of Coxsackie virus (A16), enteric cytopathic human orphan virus (ECHO type30), enterovirus (EV71), rotavirus (LLR strain), and enteric adenovirus (AdV 41), specificity assessment demonstrated that there was no cross reaction during the usage of the two test strips as shown by the results which were negative. In the detection of influenza A in 1085 clinical specimens, the total coincidence rate was 96.7%, the positive coincidence rate was 97.1%, and the negative coincidence rate was 96.7%. In the case of influenza B detection, the total coincidence rate was 99.1%, the positive coincidence rate was 92.6%, and the negative coincidence rate was 98.5%. In addition, with influenza A or B real-time RT-PCR detection method, the results showed that, for influenza A, 26 of the 33 specimens that negative with CMT but positive with FMT, showed positive results, and none of the 3 specimens that positive with CMT but negative with FMT showed a positive result; For influenza B, 12 of the 15 specimens that negative with CMT but positive with FMT, showed positive results, and none of the 5 specimens that positive with CMT but negative with FMT showed a positive result. Conclusions FMT performed better than CMT in the combined detection of influenza A and B viruses.

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A compact, low-cost, quantitative and multiplexed fluorescence detection platform for point-of-care applications

Cited by 43 publications

References 20 publications

Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform

Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform

A low-cost fluorescence reader for in vitro transcription and nucleic acid detection with Cas13a

Comparison of the detection performance of two different one-step-combined test strips with fluorescent microspheres or colored microspheres as tracers for influenza A and B viruses

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