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Development of a lateral flow immunoassay for rapid field detection of the red imported fire ant, Solenopsis invicta (Hymenoptera: Formicidae)


The red imported fire ant, Solenopsis invicta, is an aggressive, highly invasive pest ant species from South America that has been introduced into North America, Asia, and Australia. Quarantine efforts have been imposed in the USA to minimize further spread of the ant. To aid the quarantine efforts, there remains an acute need for a rapid, field portable method for the identification of these ants. In this report, we describe two novel monoclonal antibodies that specifically bind the S. invicta venom protein 2 produced by S. invicta. Using these monoclonal antibodies we developed a lateral flow immunoassay that provides a rapid and portable method for the identification of S. invicta ants. The lateral flow immunoassay was validated against purified S. invicta venom protein 2 and 33 unique ant species (representing 15 % of the total species and 42 % of the Myrmicinae genera found in Florida), and only S. invicta and the S. invicta/richteri hybrid produced a positive result. These monoclonal antibodies were selective to S. invicta venom protein 2 and did not bind to proteins from congeners (i.e., S. geminata or S. richteri) known to produce a S. invicta venom protein 2 ortholog. This S. invicta lateral flow immunoassay provides a new tool for regulatory agencies in the USA to enforce quarantine protocols and limit the spread of this invasive ant.

Field method to detect and identify the red imported fire ant, Solenopsis invicta

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We thank D. Oi (USDA-ARS, Gainesville, FL) and R. Hnasko (USDA-ARS, Albany, CA) for critical reviews of the manuscript and R. Hnasko for valuable discussions on antibodies and LFA technique. We also thank Dr. J. Oliver (Tennessee State University) for providing the Solenopsis richteri samples. The use of trade, firm, or corporation names in this publication is for the information and convenience of the reader. Such use does not constitute an official endorsement or approval by the United States Department of Agriculture or the Agricultural Research Service of any product or service to the exclusion of others that may be suitable. This material was made possible, in part, by a Cooperative Agreement from the United States Department of Agriculture’s Animal and Plant Health Inspection Service (APHIS) and may not necessarily express the views of APHIS.

Author contributions

Conceived and designed the experiments: S.M. Valles. Performed the experiments: S.M. Valles, C.A. Strong. Analyzed the data: S. M. Valles, C.A. Strong. Wrote the paper: S. M. Valles, A-M.A Callcott.

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Correspondence to Steven M. Valles.

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Antibody production was completed under contract by ProMab Biotechnologies in compliance with state and federal laws.

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The authors declare no conflicts of interest.

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Valles, S.M., Strong, C.A. & Callcott, AM.A. Development of a lateral flow immunoassay for rapid field detection of the red imported fire ant, Solenopsis invicta (Hymenoptera: Formicidae). Anal Bioanal Chem 408, 4693–4703 (2016).

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  • Solenopsis invicta
  • Solenopsis richteri
  • Fire ant
  • Venom
  • Antibodies
  • Lateral flow immunoassay