Plastic enzyme-linked immunosorbent assays (ELISA)-on-a-chip biosensor for botulinum neurotoxin A

Han, Seung-Mok; Cho, Joung-Hwan; Cho, Il-Hoon; Paek, Eui-Hwan; Oh, Hee-Bok; Kim, Bongsu; Ryu, Chunsun; Lee, Kyung‐Hee; Kim, Young-Kee; Paek, Se‐Hwan

doi:10.1016/j.aca.2007.01.028

Cited by 55 publications

(18 citation statements)

References 23 publications

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“…A system using a cross-flow immunochromatographic method was developed to enhance the sensitivity of the lateral flow BoNT assay [93]. This assay relies upon a detection antibody conjugated to HRP, instead of the more common gold nanoparticles or colloidal gold reporters.…”

Section: Lateral Flow Testsmentioning

confidence: 99%

“…The HRP chromogenic substrate migrates over the HRP enzyme in the capture zone and its cleavage translates into a color change, signaling presence of the toxin. The sensitivity of this assay is 2 ng/mL (200 MLD 50 /ml) of BoNT/A [93]; a modest enhancement compared to gold nanoparticle-based devices is paid for by increased complexity of the device and the assay protocol. A more effective and simpler signal enhancement procedure using silver enhancer has been developed [94].…”

Section: Lateral Flow Testsmentioning

confidence: 99%

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Sensing the Deadliest Toxin: Technologies for Botulinum Neurotoxin Detection

Čapek

Dickerson

2010

Toxins

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Sensitive and rapid detection of botulinum neurotoxins (BoNTs), the most poisonous substances known to date, is essential for studies of medical applications of BoNTs and detection of poisoned food, as well as for response to potential bioterrorist threats. Currently, the most common method of BoNT detection is the mouse bioassay. While this assay is sensitive, it is slow, quite expensive, has limited throughput and requires sacrificing animals. Herein, we discuss and compare recently developed alternative in vitro detection methods and assess their ability to supplement or replace the mouse bioassay in the analysis of complex matrix samples.

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Section: Lateral Flow Testsmentioning

confidence: 99%

Section: Lateral Flow Testsmentioning

confidence: 99%

Sensing the Deadliest Toxin: Technologies for Botulinum Neurotoxin Detection

Čapek

Dickerson

2010

Toxins

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“…This 1.25 nM LOD is equivalent to 62.5 ng/ml of the LcA and is comparable to similar studies by Schmidt and Stafford (Schmidt and Stafford, 2003) using a SNAP-25 peptide substrate labeled with a different FRET pair (1 nM; 60 ng/ml), measured using a standard fluorimeter. When compared to a number of immunoassay formats including ELISAs (Han et al, 2007), some commercial lateral flow test kits (Gessler et al, 2007) and fluorescent immunoassays the LOD is in the same nanogram per milliliter range (2-50 ng/ml). Some more sophisticated detection schemes, such as those combining paramagnetic beads and electrochemiluminescent immunoassays, have measured down to 50 pg/ml of BoTN-A (Rivera et al, 2006).…”

Section: El-ccd Point Of Care Detection Of Botulinum Toxin Activitymentioning

confidence: 99%

“…The mouse bioassay is very sensitive and can detect as little as 0.03 ng of botulinum toxin (1 ng = 30 mouse 50% lethal doses) within 6-96 h Johnson, 1990, 1992). Several different immunoassays have been developed for BoTN-A detection including, enzyme-linked immunosorbent assays (ELISA) (Ferreira et al, 2004;Guglielmo-Viret et al, 2005;Han et al, 2007;Keller et al, 1999;Peruski et al, 2002;Sharma et al, 2006), lateral flow (Gessler et al, 2007;Sharma et al, 2005), immunomagnetic beads (Gessler et al, 2006;Rivera et al, 2006), immunoaffinity column (Attree et al, 2007;Gessler et al, 2005) and immunosensors (Golden et al, 2005;Ligler et al, 2007;Ligler et al, 2003). While these immunoassays have similar sensitivities to the mouse bioassay, and are much more rapid, they do not provide information about the activity state of the toxin.…”

Section: Introductionmentioning

confidence: 99%

A fluorescence detection platform using spatial electroluminescent excitation for measuring botulinum neurotoxin A activity

Sapsford

Sun

Francis

et al. 2008

Biosensors and Bioelectronics

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Current biodetection illumination technologies (laser, LED, tungsten lamp, etc.) are based on spot illumination with additional optics required when spatial excitation is required. Herein we describe a new approach of spatial illumination based on electroluminescence (EL) semiconductor strips available in several wavelengths, greatly simplifying the biosensor design by eliminating the need for additional optics. This work combines EL excitation with charge-coupled device (CCD) based detection (EL-CCD detector) of fluorescence for developing a simple portable detector for botulinum neurotoxin A (BoTN-A) activity analysis. A Förster Resonance Energy Transfer (FRET) activity assay for BoTN-A was used to both characterize and optimize the EL-CCD detector. The system consists of two modules: (1) the detection module which houses the CCD camera and emission filters, and (2) the excitation and sample module, containing the EL strip, the excitation filter and the 9-well sample chip. The FRET activity assay used in this study utilized a FITC/DABCYL-SNAP-25 peptide substrate in which cleavage of the substrate by BoTN-A, or its light chain derivative (LcA), produced an increase in fluorescence emission. EL-CCD detector measured limits of detection (LODs) were similar to those measured using a standard fluorescent plate reader with valves between 0.625 and 1.25 nM (31-62 ng/ml) for LcA and 0.313 nM (45 ng/ml) for the full toxin, BoTN-A. As far as the authors are aware this is the first demonstration of phosphor-based EL strips being used for the spatial illumination/excitation of a surface, coupled with CCD for point of care detection.

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“…The antibody-antigen complex further continues to migrate along the strip and is captured in the detection zone by a capture antibody, resulting in a visible change of color. Several LFAs for the detection of different BoNT serotypes have been developed with detection limits usually between 0.3 and 250 ng/mL (Chiao et al 2004(Chiao et al , 2008aKlewitz et al 2006;Attrée et al 2007;Han et al 2007). Sharma and co-workers evaluated two commercial products on spiked food samples and found detection limits of above 20 ng/mL for BoNT complexes of serotypes A, B, and E (Sharma et al 2005).…”

Section: Rapid Detection Tools Based On Elisa Formatsmentioning

confidence: 99%

Complexity of Botulinum Neurotoxins: Challenges for Detection Technology

Dorner

Schulz

Kull

et al. 2012

Current Topics in Microbiology and Immunology

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The detection of botulinum neurotoxins (BoNT) is extremely challenging due to their high toxicity and the multiple BoNT variants. To date, seven serotypes with more than 30 subtypes have been described, and even more subtypes are expected to be discovered. The fact that the BoNT molecules are released as large complexes of different size and composition adds further complexity to the issue. Currently, in the diagnostics of botulism, the mouse bioassay (MBA) is still considered as gold standard for the detection of BoNT in complex sample materials. Over the years, different functional, immunological, and spectrometric assays or combinations thereof have been developed, supplemented by DNA-based assays for the detection of the organism. In this review, advantages and limitations of the current technologies will be discussed, highlighting some of the intricacies of real sample analysis.

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Plastic enzyme-linked immunosorbent assays (ELISA)-on-a-chip biosensor for botulinum neurotoxin A

Cited by 55 publications

References 23 publications

Sensing the Deadliest Toxin: Technologies for Botulinum Neurotoxin Detection

Sensing the Deadliest Toxin: Technologies for Botulinum Neurotoxin Detection

A fluorescence detection platform using spatial electroluminescent excitation for measuring botulinum neurotoxin A activity

Complexity of Botulinum Neurotoxins: Challenges for Detection Technology

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