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Diseases of Aquatic Organisms

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DAO 60:233-240 (2004)  -  doi:10.3354/dao060233

Development of monoclonal antibodies fordetection of necrotizing hepatopancreatitisin penaeid shrimp

Deborah J. Bradley-Dunlop*, Carlos Pantoja, Donald V. Lightner

Department of Veterinary Science and Microbiology, University of Arizona, Tucson, Arizona 85721, USA

ABSTRACT: Monoclonal antibodies (MAbs) were produced against necrotizing hepatopancreatitis bacteria (NHP-B) of penaeid shrimp. The MAbs tested in dot-immunoblot (D-IB) assays were capable of detecting the NHP-B in hepatopancreas samples collected from moribund juvenile Litopenaeus vannamei during an experimentally induced NHP-B infection. The MAbs were also screened by immunohistochemistry (IHC) using case submissions that were determined to be infected not only by histology, but also polymerase chain reaction (PCR) and in situ hybridization (ISH) assays using specific digoxigenin (DIG)-labeled probes on histological sections prepared from naturally infected shrimp. Two of the MAbs were chosen for development of detection methods for NHP. The MAbs were tested using IHC methods on Davidson’s alcohol-formalin-acetic acid (AFA) fixed tissue sections and identified NHP-B infected cells and tissues in a pattern similar to that seen with DIG-labeled NHP-specific gene probes. None of the MAbs reacted with tissue from specific pathogen-free (SPF) shrimp or with shrimp tissues infected with a rickettsia-like bacteria, Vibrio sp., Campylobacter sp., and Spiroplasma sp. The MAbs were found to be negative against these other organisms, demonstrating that they are species specific and useful for rapid diagnostic detection of NHP-B.

KEY WORDS: NHP · Necrotizing hepatopanceatitis · Monoclonal antibodies · Dot-immunoblot · Immunohistochemistry

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