Direct Competitive Enzyme-Linked Immunosorbent Assay for Saxitoxin and Neosaxitoxin

Huang, Xuan; Hsu, Kuo-Hui; Chu, Fun Sun

doi:10.1021/jf950717j

Cited by 24 publications

(20 citation statements)

References 30 publications

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“…Since then, additional indirect assays have been developed for STX and neoSTX, with a detection limit of 10 pg/mL [105,106]. Direct assays have also been developed for STX, neoSTX and GTX2/3, with detection limits in the pg/mL range [106,107,108,109]. …”

Section: Methods Of Detectionmentioning

confidence: 99%

An Overview on the Marine Neurotoxin, Saxitoxin: Genetics, Molecular Targets, Methods of Detection and Ecological Functions

2013

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Marine neurotoxins are natural products produced by phytoplankton and select species of invertebrates and fish. These compounds interact with voltage-gated sodium, potassium and calcium channels and modulate the flux of these ions into various cell types. This review provides a summary of marine neurotoxins, including their structures, molecular targets and pharmacologies. Saxitoxin and its derivatives, collectively referred to as paralytic shellfish toxins (PSTs), are unique among neurotoxins in that they are found in both marine and freshwater environments by organisms inhabiting two kingdoms of life. Prokaryotic cyanobacteria are responsible for PST production in freshwater systems, while eukaryotic dinoflagellates are the main producers in marine waters. Bioaccumulation by filter-feeding bivalves and fish and subsequent transfer through the food web results in the potentially fatal human illnesses, paralytic shellfish poisoning and saxitoxin pufferfish poisoning. These illnesses are a result of saxitoxin’s ability to bind to the voltage-gated sodium channel, blocking the passage of nerve impulses and leading to death via respiratory paralysis. Recent advances in saxitoxin research are discussed, including the molecular biology of toxin synthesis, new protein targets, association with metal-binding motifs and methods of detection. The eco-evolutionary role(s) PSTs may serve for phytoplankton species that produce them are also discussed.

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Section: Methods Of Detectionmentioning

confidence: 99%

An Overview on the Marine Neurotoxin, Saxitoxin: Genetics, Molecular Targets, Methods of Detection and Ecological Functions

2013

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“…The PST levels of the samples are presented in terms of the amount of STX, equivalent to the total toxicity exerted by a mixture of several different PSTs contained in the samples. As shown in Figure 6, the absorbance was inversely related to the total PST level in the shellfish samples, with an r 2 value of 0.96, even though different PSTs in the samples have different reactivity towards each anti-STX antibody 7,13,25 . Moreover, the current regulatory limit (80 μg of STX equivalents/100 g shellfish meat) was within the dynamic range of the method.…”

Section: Pst Assaymentioning

confidence: 94%

“…Other techniques, including HPLC 4-6 , ELISA [7][8][9][10][11][12] , and immunoassay methods coupled with other detection strategies (such as surface plasmon resonance 13,14 , polymerase chain reaction 15 , microsphereflow cytometry 16,17 , and microarray 18,19 ) are unsuitable for field applications because they require instrumentation that is confined to the lab environment. Although lateral flow immunoassay methods have been developed for field monitoring 20,21 , they suffer from low sensitivity and reliability 22 .…”

Section: Introductionmentioning

confidence: 99%

Development of an improved stationary liquid-phase lab-on-a-chip for the field monitoring of paralytic shellfish toxins

Choi

2016

BioChip J

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A stationary liquid-phase lab-on-a-chip (SLP-LOC), in which magnetic particles are used as a solid phase, has been previously reported as a novel platform for immunoassay methods. The SLP-LOC had a simple structure with two chambers connected by a narrow channel. The assay was performed by transporting the particles from one chamber to the next while the liquid phases were in a stationary state. However, each chamber was frequently contaminated by other chamber materials because of the small volume of the channel. Consequently, in this work, we have developed a three-chamber SLP-LOC (3C-LOC) featuring an additional chamber containing a buffered washing solution positioned between the existing two chambers and connected by channels. The dimensions of the 3C-LOC were optimized to fit into a standard spectrophotometer cuvette holder, allowing the direct measurement of absorbance. The 3C-LOC exhibited improved washing efficiency, reliability, and sensitivity in an assay of standard saxitoxin samples. Furthermore, the method was used to effectively determine whether paralytic shellfish toxin (PST) levels in shellfish samples exceeded regulatory limits. The results of this study indicate that the 3C-LOC could provide a simple and rapid method to monitor PSTs in the field.

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“…To simplify this analysis, Huang et al [ 89 ] used two antibodies within a single heterologous ELISA to produce an assay capable of detecting both STX and NEO at the same time. By combining two antibodies, Huang et al [ 89 ] were able to increase the sensitivity and specificity of their test to detect total PSP levels similar to the MBA.…”

Section: Methods For End-product Testing (Ept)mentioning

confidence: 99%

Current Trends and Challenges for Rapid SMART Diagnostics at Point-of-Site Testing for Marine Toxins

Dillon

Zaczek-Moczydlowska

Edwards

et al. 2021

Sensors

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In the past twenty years marine biotoxin analysis in routine regulatory monitoring has advanced significantly in Europe (EU) and other regions from the use of the mouse bioassay (MBA) towards the high-end analytical techniques such as high-performance liquid chromatography (HPLC) with tandem mass spectrometry (MS). Previously, acceptance of these advanced methods, in progressing away from the MBA, was hindered by a lack of commercial certified analytical standards for method development and validation. This has now been addressed whereby the availability of a wide range of analytical standards from several companies in the EU, North America and Asia has enhanced the development and validation of methods to the required regulatory standards. However, the cost of the high-end analytical equipment, lengthy procedures and the need for qualified personnel to perform analysis can still be a challenge for routine monitoring laboratories. In developing regions, aquaculture production is increasing and alternative inexpensive Sensitive, Measurable, Accurate and Real-Time (SMART) rapid point-of-site testing (POST) methods suitable for novice end users that can be validated and internationally accepted remain an objective for both regulators and the industry. The range of commercial testing kits on the market for marine toxin analysis remains limited and even more so those meeting the requirements for use in regulatory control. Individual assays include enzyme-linked immunosorbent assays (ELISA) and lateral flow membrane-based immunoassays (LFIA) for EU-regulated toxins, such as okadaic acid (OA) and dinophysistoxins (DTXs), saxitoxin (STX) and its analogues and domoic acid (DA) in the form of three separate tests offering varying costs and benefits for the industry. It can be observed from the literature that not only are developments and improvements ongoing for these assays, but there are also novel assays being developed using upcoming state-of-the-art biosensor technology. This review focuses on both currently available methods and recent advances in innovative methods for marine biotoxin testing and the end-user practicalities that need to be observed. Furthermore, it highlights trends that are influencing assay developments such as multiplexing capabilities and rapid POST, indicating potential detection methods that will shape the future market.

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Direct Competitive Enzyme-Linked Immunosorbent Assay for Saxitoxin and Neosaxitoxin

Cited by 24 publications

References 30 publications

An Overview on the Marine Neurotoxin, Saxitoxin: Genetics, Molecular Targets, Methods of Detection and Ecological Functions

An Overview on the Marine Neurotoxin, Saxitoxin: Genetics, Molecular Targets, Methods of Detection and Ecological Functions

Development of an improved stationary liquid-phase lab-on-a-chip for the field monitoring of paralytic shellfish toxins

Current Trends and Challenges for Rapid SMART Diagnostics at Point-of-Site Testing for Marine Toxins

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