Paralytic shellfish toxins in clinical matrices: Extension of AOAC official method 2005.06 to human urine and serum and application to a 2007 case study in Maine

DeGrasse, Stacey L.; Rivera, Victor R.; Roach, John A. G.; White, Kevin D.; Callahan, John H.; Couture, Darcie A.; Simone, Karen; Peredy, Tamas R.; Poli, Mark

doi:10.1016/j.dsr2.2012.08.001

Cited by 24 publications

(20 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…in water samples that also test positive for DA with the ASP Jellett Rapid Testing kit. Most offshore U.S. waters, including Georges Bank, fall under federal jurisdiction and are managed by the U.S. Food and Drug Administration, primarily for PSP toxins at this time (DeGrasse et al, this issue). Other states, including those along the U.S. west coast where DA closures occur regularly (Lewitus et al, 2012; Trainer et al, 2009a; Trainer et al, 2009b), conduct routine shellfish flesh testing for DA.…”

Section: Introductionmentioning

confidence: 99%

Diversity and toxicity of the diatom Pseudo-nitzschia Peragallo in the Gulf of Maine, Northwestern Atlantic Ocean

Fernandes

Hubbard

Richlen

et al. 2014

Deep Sea Research Part II: Topical Studies in Oceanography

View full text Add to dashboard Cite

Multiple species in the toxic marine diatom genus Pseudo-nitzschia have been identified in the Northwestern Atlantic region encompassing the Gulf of Maine (GOM), including the Bay of Fundy (BOF). To gain further knowledge of the taxonomic composition and toxicity of species in this region, Pseudo-nitzschia isolates (n=146) were isolated from samples collected during research cruises that provided broad spatial coverage across the GOM and the southern New England shelf, herein referred to as the GOM region, during 2007-2008. Isolates, and cells in field material collected at 38 stations, were identified using electron microscopy (EM). Eight species (P. americana, P. fraudulenta, P. subpacifica, P. heimii, P. pungens, P. seriata, P. delicatissima and P. turgidula), and a novel form, Pseudo-nitzschia sp. GOM, were identified. Species identity was confirmed by sequencing the large subunit of the ribosomal rDNA (28S) and the internal transcribed spacer 2 (ITS2) for six species (36 isolates). Phylogenetic analyses (including neighbor joining, maximum parsimony, and maximum likelihood estimates and ITS2 secondary structure analysis) and morphometric data supported the placement of P. sp. GOM in a novel clade that includes morphologically and genetically similar isolates from Australia and Spain and is genetically most similar to P. pseudodelicatissima and P. cuspidata. Seven species (46 isolates) were grown in nutrient-replete batch culture and aliquots consisting of cells and growth medium were screened by Biosense ASP ELISA to measure total domoic acid (DA) produced (intracellular + extracellular); P. americana and P. heimii were excluded from all toxin analyses as they did not persist in culture long enough for testing. All 46 isolates screened produced DA in culture and total DA varied among species (e.g., 0.04 to 320 ng ml-1 for P. pungens and P. sp. GOM isolates, respectively) and among isolates of the same species (e.g., 0.24 – 320 ng ml-1 for P. sp. GOM). The 15 most toxic isolates corresponded to P. seriata, P. sp. GOM and P. pungens, and fg DA cell-1 was determined for whole cultures (cells and medium) using ELISA and liquid chromatography (LC) with fluorescence detection (FLD); for seven isolates, toxin levels were also estimated using LC - with mass spectrometry and ultraviolet absorbance detection. Pseudo-nitzschia seriata was the most toxic species (up to 3,500 fg cell-1) and was observed in the GOM region during all cruises (i.e., during the months of April, May, June and October). Pseudo-nitzschia sp. GOM, observed only during September and October 2007, was less toxic (19 – 380 fg cell-1) than P. seriata but more toxic than P. pungens var. pungens (0. 4 fg cell-1). Quantitation of DA indicated that concentrations measured by LC and ELISA were positively and significantly correlated; the lower detection limit of the ELISA permitted quantification of toxicity in isolates that were found to be nontoxic with LC methods. The confirmation of at least seven toxic species and the broad spatial and temporal dis...

show abstract

Section: Introductionmentioning

confidence: 99%

Diversity and toxicity of the diatom Pseudo-nitzschia Peragallo in the Gulf of Maine, Northwestern Atlantic Ocean

Fernandes

Hubbard

Richlen

et al. 2014

Deep Sea Research Part II: Topical Studies in Oceanography

View full text Add to dashboard Cite

show abstract

“…To our knowledge, this is the first validated method for quantitation of STX in whole blood. When compared with analytical methods for detection of STX in other clinical matrices, this method is approximately 50 times more sensitive in whole blood and more than an order of magnitude more sensitive in dried blood than currently developed LC/MS-MS methods for quantitation of STX in urine and approximately 200 times more sensitive in whole blood and 66 times more sensitive in dried blood than previously reported LC-MS methods for serum (Bragg et al, 2015; DeGrasse et al, 2014; Johnson et al, 2009). Because of this method’s high sensitivity compared with other techniques, it would be advantageous to add as a screening measure of PSP exposures so as not to report out false negatives.…”

Section: Discussionmentioning

confidence: 92%

“…There is currently very little published data on STX levels in clinical specimens. Two reports of STX exposures identified STX levels of up to 51 nM (15.3 ng/mL) in serum and 3.4 μM (1017 ng/mL) in urine during acute exposure (DeGrasse et al, 2014; Gessner et al, 1997). Because onset of symptoms occur quickly (within 1–2 h of exposure) and the half-life of STX is short (less than 10 h in serum and 20.4 h in urine), it is important to develop methods that are rapid, accurate, and sensitive to detect STX in clinical samples and confirm diagnosis.…”

Section: Discussionmentioning

confidence: 99%

“…Despite monitoring programs which ban harvesting seafood with an excess of 80 μg STX equivalents/100 g of tissue, there are an estimated 2000 worldwide human cases of PSP reported yearly, with a mortality rate of 15% (Etheridge, 2010; Van Dolah, 2000). Limited case reports suggest that the half-life of STX is less than 10 h in serum and 20.4 h in urine (DeGrasse et al, 2014; Gessner et al, 1997). Because of its potency and threat to human health along with its short-half life in vivo , there is a need for rapid and sensitive diagnostic techniques to monitor human exposures and confirm diagnoses.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Quantification of saxitoxin in human blood by ELISA

Wharton

Feyereisen

Gonzalez

et al. 2017

Toxicon

View full text Add to dashboard Cite

Saxitoxin (STX) is a potent marine toxin that causes paralytic shellfish poisoning (PSP) which can result in significant morbidity and mortality in humans. Low lethal doses, rapid onset of PSP symptoms, and brief STX half-life in vivo require sensitive and rapid diagnostic techniques to monitor human exposures. Our laboratory has validated an enzyme-linked immunosorbent assay (ELISA) for quantitative detection of STX from 0.020 to 0.80 ng/mL in human whole blood and from 0.06 to 2.0 ng/mL in dried human blood which is simple, sensitive, rapid, and cost-effective. To our knowledge, this is the first validated method for the quantitation of saxitoxin in whole blood. Microsampling devices were used in sample collection which allows for standardized collection of blood, stable storage, and cost-efficient shipping. Quality control precision and accuracy were evaluated over the course of validation and were within 20% of theoretical concentrations. No detectable background concentrations of STX were found among fifty whole blood and dried blood convenience samples. Additionally, ten spiked individual whole blood and dried blood samples were tested for accuracy and precision and were within 20% of theoretical concentrations. Gonyautoxins 2&3 (GTX2&3) cross-reacted with this ELISA by 21%, but all other structurally related PSP toxins tested cross-reacted less than two percent. While clinical diagnosis or treatment of PSP would be unaffected by GTX2&3 cross-reactivity by ELISA, to accurately quantify individual PSP toxins, these results should be coupled with high performance liquid chromatography mass spectrometry measurements.

show abstract

“…Along this line, recently the official pre-column method (AOAC 2005.06) was extended to human urine and serum [80]. The complexity of clinical matrices and the current lack of standard operating procedures for clinical samples, however, make the identification and quantification of the PSP toxins rather difficult.…”

Section: Review and Discussion Of Trends In Analytical Methods For Bimentioning

confidence: 98%

Biological toxins of potential bioterrorism risk: Current status of detection and identification technology

Dorner

Zeleny

Harju

et al. 2016

TrAC Trends in Analytical Chemistry

View full text Add to dashboard Cite

Paralytic shellfish toxins in clinical matrices: Extension of AOAC official method 2005.06 to human urine and serum and application to a 2007 case study in Maine

Cited by 24 publications

References 18 publications

Diversity and toxicity of the diatom Pseudo-nitzschia Peragallo in the Gulf of Maine, Northwestern Atlantic Ocean

Diversity and toxicity of the diatom Pseudo-nitzschia Peragallo in the Gulf of Maine, Northwestern Atlantic Ocean

Quantification of saxitoxin in human blood by ELISA

Biological toxins of potential bioterrorism risk: Current status of detection and identification technology

Contact Info

Product

Resources

About