Multiplexed lateral flow immunoassay to discriminate Solenopsis invicta, Solenopsis richteri, and Solenopsis invicta × richteri hybrids
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Solenopsis invicta and Solenopsis richteri are aggressive, highly invasive ant species from South America that were introduced into North America in the early part of the twentieth century. Biosecurity efforts in the US included the establishment of a quarantine to minimize the human-assisted spread of these ants. A limitation of the quarantine is rapid identification/discrimination of these ants when found entering non-quarantined areas. Using monoclonal antibodies designed toward S. invicta and S. richteri venom protein 2, we developed a multiplexed lateral flow immunoassay that provides a rapid and portable method for the identification of S. invicta, S. richteri, and the S. invicta × richteri hybrid. The multiplexed lateral flow immunoassay was validated against 39 unique ant species, and only S. invicta, S. richteri, and the S. invicta × richteri hybrid were detected. The assay did not detect proteins from the congener S. geminata known to produce a S. invicta venom protein 2 ortholog. The invasive fire ant multiplexed lateral flow immunoassay provides a new tool for regulatory agencies in the US to enforce quarantine protocols and limit the spread of these invasive ants.
KeywordsSolenopsis invicta Solenopsis richteri Fire ant Venom Antibodies Lateral flow immunoassay
We thank R. Hnasko (USDA-ARS, Albany, CA) for valuable discussions on antibodies and the LFA technique, Drs. J. Oliver (Tennessee State University), and J. Chen (USDA-ARS, Stoneville, MS) for providing samples of Solenopsis richteri and S. invicta × richteri hybrid samples and Dr. K. Addesso (Tennessee State University) for conducting the cuticular hydrocarbon and venom alkaloid analyses on the hybrid fire ants. 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 APHIS’ views.
Conceived and designed the experiments: SMV. Performed the experiments: SMV and CAS. Analyzed the data: SMV and CAS. Wrote the paper: SMV and A-MAC.
Compliance with ethical standards
The authors ensure that this research has been carried out in accordance with the “International Guiding Principles for Biomedical Research Involving Animals,” as revised by the International Council for Laboratory Animal Science (ICLAS) and the Councils for International Organizations of Medical Sciences (CIOMS) in 2012, and that any research involving live vertebrates performed by scientists at the USDA Agricultural Research Service was reviewed and approved by a properly constituted IACUC as required in ARS Policies and Procedures Manual 130.4, version 2. The antibodies used in this project were purchased from a commercial vendor (ProMab Biotechnologies, Richmond, CA) that is not subject to the oversight authority of ARS.
Conflict of interest
The authors declare no conflicts of interest.
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