Development of monoclonal antibody-based assays for the detection of Vibrio tubiashii zinc-metalloprotease (VtpA)

Gharaibeh, Dima N.; Biel, Frances M.; Häse, Claudia C.

Citeable URL: https://ir.library.oregonstate.edu/concern/articles/wd375x038

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Descriptions

Attribute Name	Values
Creator	Gharaibeh, Dima N. Biel, Frances M. Häse, Claudia C.
Abstract	Vibrio tubiashii has been linked to disease outbreaks in molluscan species, including oysters, geoducks, and clams. In particular, oyster hatcheries in the Pacific Northwest have been plagued by intermittent vibriosis since 2006. Accurate detection of vibrios, including V. tubiashii, is critical to the hatcheries in order to allow for rapid remediation efforts. The current methods for detection of Vibrio spp. are not ideal for use at the hatchery. Plating samples require time and is not sensitive to extracelluar pathogenic products, such as the secreted zinc-metalloprotease, VtpA. Other sensitive methods to detect bacteria, such as qPCR, require a high level of laboratory skills and expensive supplies that are prohibitive for use at hatchery sites. Thus, hatcheries would benefit from a sensitive, simple method to detect V. tubiashii and its secreted toxin. Here, we describe the development of two inexpensive and highly specific tests for the shellfish—toxic zinc-metalloprotease secreted by V. tubiashii: enzyme-linked immunoassays (ELISA) and a lateral flow immunoassay (dipstick assay). Both technologies rely on a set of monoclonal antibodies used in a sandwich format, with the capture antibody recognizing a different epitope than the detection antibody on the mature VtpA protein. Both assays are quantitative and give colorimetric readouts. The sandwich ELISA was sensitive when VtpA was diluted into PBS, but was markedly less sensitive in conditions that correlate with the environment of hatchery-derived samples, such as in the presence of seawater, algae, or oyster larvae. In contrast, the dipstick assay remained very sensitive in the presence of these contaminants, is less work-intensive, and much more rapid, making this format the preferred assay method for detecting VtpA on site in a hatchery or environmental setting.
License	All rights reserved
Resource Type	Article
DOI	https://doi.org/10.1016/j.mimet.2013.05.009
Date Available	2013-05-17T20:52:36+00:00
Date Issued	2013-05-09
Citation	Gharaibeh, D. N., Biel, F. M., & Häse, C. C. (2013). Development of monoclonal antibody-based assays for the detection of vibrio tubiashii zinc-metalloprotease (VtpA). Journal of Microbiological Methods, 94(2), 125-132. doi:10.1016/j.mimet.2013.05.009
Journal Title	Journal of Microbiological Methods
Journal Volume	94
Journal Issue/Number	2
Academic Affiliation	College of Veterinary Medicine
Rights Statement	In Copyright
Funding Statement (additional comments about funding)	This research was supported by grantsfrom the USDA (award numbers ORE00246, ORE00088) and NOAA (contract numberNA09NMF4270088).
Publisher	Elsevier
Peer Reviewed	Yes
Language	English [eng]
Replaces	http://hdl.handle.net/1957/38608
Additional Information	description.provenance : Approved for entry into archive by Sue Kunda(sue.kunda@oregonstate.edu) on 2013-05-17T20:52:36Z (GMT) No. of bitstreams: 1 HaseClaudiaVetMedDevelopmentMonoclonalAntibodyBased.pdf: 468213 bytes, checksum: 8810af9454a067c057fcaef7f1661afa (MD5) description.provenance : Submitted by Sue Kunda (sue.kunda@oregonstate.edu) on 2013-05-17T20:51:43Z No. of bitstreams: 1 HaseClaudiaVetMedDevelopmentMonoclonalAntibodyBased.pdf: 468213 bytes, checksum: 8810af9454a067c057fcaef7f1661afa (MD5) description.provenance : Made available in DSpace on 2013-05-17T20:52:36Z (GMT). No. of bitstreams: 1 HaseClaudiaVetMedDevelopmentMonoclonalAntibodyBased.pdf: 468213 bytes, checksum: 8810af9454a067c057fcaef7f1661afa (MD5) Previous issue date: 2013-05-09

ScholarsArchive@OSU

Article