研究目的
To develop a simple, inexpensive, and portable smartphone-based fluorescent lateral flow immunoassay platform for highly sensitive point-of-care detection of Zika virus nonstructural protein 1 (ZIKV NS1) in low-resource settings.
研究成果
The smartphone-based fluorescent LFIA platform enables rapid, quantitative detection of ZIKV NS1 with high sensitivity and specificity, showing low cross-reactivity and good performance in complex biological samples. It is portable, low-cost, and suitable for point-of-care applications in resource-limited settings, offering a promising tool for early diagnosis of Zika virus infections.
研究不足
The assay requires a four-fold dilution of serum samples to reduce matrix effects, which may affect sensitivity. The device has a limited battery life of ~2 hours, and the cost could be further reduced by removing filters, but this might increase background noise. Cross-reactivity with other flaviviruses, although limited, could still be a concern in endemic areas.
1:Experimental Design and Method Selection:
The study involved designing a smartphone-based fluorescent LFIA reader using a 3D-printed attachment to integrate optical components. Quantum dot microspheres were chosen as fluorescent probes for their bright signals.
2:Sample Selection and Data Sources:
ZIKV NS1 protein, antibodies, and DENV NS1 were purchased; inactivated ZIKV virions and normal human sera were provided or collected.
3:List of Experimental Equipment and Materials:
Included a smartphone (Nokia Lumia 1020), UV LED (5 W, SVC), optical filters (Edmund Optical), lenses, power unit with lithium battery (Sony), QD microspheres (Najing Technology), NC membranes (Millipore), and various pads and cassettes (Jieyi Biotechnology).
4:Experimental Procedures and Operational Workflow:
Conjugation of QD microspheres with antibodies, preparation of LFIA strips, detection of ZIKV NS1 in buffer and serum samples by adding sample to strips, incubating for 20 min, and imaging with the smartphone device.
5:Data Analysis Methods:
Fluorescence images were analyzed using ImageJ software with Gel Analyzer plug-in to calculate test line intensities.
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Short-pass filter
425 nm
Edmund Optical
Filters the UV light to reduce background noise by blocking wavelengths above 425 nm.
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Bandpass filter
624/40 nm
Edmund Optical
Filters the fluorescence signal to allow only light around 624 nm to pass, reducing interference.
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Lens
Plano-convex lens
Edmund Optical
Reduces the object distance for the smartphone camera, improving image signal collection.
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Smartphone
Lumia 1020
Nokia
Used as the imaging and processing unit for capturing and analyzing fluorescence signals from the LFIA strips.
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UV LED
5 W
SVC
Serves as the excitation light source for the quantum dot microspheres on the LFIA strips.
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Lithium battery
3000 mAh, 3.7 V
Sony
Powers the UV LED and other electrical components in the device.
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QD microspheres
Carboxyl-functionalized CdSe/ZnS
Najing Technology
Used as fluorescent probes in the LFIA for detecting ZIKV NS1, providing bright signals.
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NC membrane
Hi-Flow Plus 135
Millipore
Part of the LFIA strip where test and control lines are immobilized for antigen detection.
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ImageJ software
Used for analyzing fluorescence images to calculate test line intensities.
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