Insectes Sociaux

, Volume 65, Issue 3, pp 493–501 | Cite as

Multiplexed lateral flow immunoassay to discriminate Solenopsis invicta, Solenopsis richteri, and Solenopsis invicta × richteri hybrids

  • S. M. VallesEmail author
  • C. A. Strong
  • A.-M. A. Callcott
Research Article


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.


Solenopsis 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.

Author contributions

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

Ethical approval

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.


  1. Ascunce MS, Yang CC, Oakey J, Calcaterra L, Wu WJ, Shih CJ, Goudet J, Ross KG, Shoemaker D (2011) Global invasion history of the fire ant Solenopsis invicta. Science 331:1066–1068CrossRefPubMedGoogle Scholar
  2. Bertagnolli D, Graham LC, Vander Meer RK (2007) Ouch! Who bit me? IFA in Alabama. In: Annual imported fire ant conference, 2007, pp 112–115Google Scholar
  3. Buren WF (1972) Revisionary studies on the taxonomy of teh imorted fire ants. J Georgia Entomol Soc 7:1–26Google Scholar
  4. Callcott AM, Collins HL (1996) Invasion and range expansion of imported fire ants (Hymenoptera: Formicidae) in North America from 1918 to 1995. Fla Entomol 79:240–251CrossRefGoogle Scholar
  5. Choi S, Lee JH, Choi JS, Jung HI (2015) Economical and rapid manufacturing of lateral flow immunosensor using fountain pens and gold colloidal solution. Anal Methods UK 7:1834–1842CrossRefGoogle Scholar
  6. Deyrup M (2003) An updated list of Florida ants (Hymenoptera: Formicidae). Fla Entomol 86:43–48CrossRefGoogle Scholar
  7. Deyrup M, Cover S (2004) A new species of Odontomachus ant (Hymenoptera: Formicidae) from inland ridges of Florida, with a key to Odontomachus of the United States. Fla Entomol 87:136–144CrossRefGoogle Scholar
  8. Fisher BL, Cover SP (2007) Ants of North America, a guide to the genera. University of California Press, BerkeleyGoogle Scholar
  9. Glancey BM, Vandermeer RK, Wojcik DP (1989) Polygyny in hybrid imported fire ants. Fla Entomol 72:632–636CrossRefGoogle Scholar
  10. Haight KL, Tschinkel WR (2003) Patterns of venom synthesis and use in the fire ant, Solenopsis invicta. Toxicon 42:673–682CrossRefPubMedGoogle Scholar
  11. Hoffman DR (1995) Fire ant venom allergy. Allergy 50:535–544CrossRefPubMedGoogle Scholar
  12. Hoffman DR (1997) Reactions to less common species of fire ants. J Allergy Clin Immunol 100:679–683CrossRefPubMedGoogle Scholar
  13. Hoffman DR (2010) Ant venoms. Curr Opin Allergy Clin Immunol 10:342–346CrossRefPubMedGoogle Scholar
  14. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMedPubMedCentralGoogle Scholar
  15. MacGown JA (2003) Ants (Formicidae) of the Southeastern United States. Mississippi State University, MississippiGoogle Scholar
  16. Oliver JB, Vander Meer RK, Ochieng SA, Youssef NN, Hosack MY, Mrema FA, Vail KM, Parkman J, Pantaleoni E, Valles SM, Haun WG, Powell S (2009) Statewide survey of imported fire ant (Hymenoptera: Formicidae) populations in Tennessee. J Entomol Sci 44:149–157CrossRefGoogle Scholar
  17. Pereira RM (2003) Areawide suppression of fire ant populations in pastures: project update. J Agric Urban Entomol 20:123–130Google Scholar
  18. Porter SD, Williams DF, Patterson RS, Fowler HG (1997) Intercontinental differences in the abundance of Solenopsis fire ants (Hymenoptera: Formicidae): escape from natural enemies? Environ Entomol 26:373–384CrossRefGoogle Scholar
  19. Rabeling C, Cover SP, Johnson RA, Mueller UG (2007) A review of the North American species of the fungus-gardening ant genus Trachymyrmex (Formicidae: Hymenoptera). Zootaxa 1664:1–53Google Scholar
  20. Ross KG, Shoemaker DD (2005) Species delimitation in native South American fire ants. Mol Ecol 14:3419–3438CrossRefPubMedGoogle Scholar
  21. Ross KG, Vandermeer RK, Fletcher DJC, Vargo EL (1987) Biochemical phenotypic and genetic-studies of 2 introduced fire ants and their hybrid (Hymenoptera, Formicidae). Evolution 41:280–293CrossRefPubMedGoogle Scholar
  22. Shoemaker DD, Ross KG, Arnold ML (1996) Genetic structure and evolution of a fire ant hybrid zone. Evolution 50:1958–1976CrossRefPubMedGoogle Scholar
  23. Streett DA, Freeland TB, Vander Meer RK (2006) Survey of imported fire ant (Hymenoptera: Formicidae) populations in Mississippi. Fla Entomol 89:91–92CrossRefGoogle Scholar
  24. Trager JC, MacGown JA, Trager MD (2007) Revision of the nearctic endemic Formica pallidefulva group. In: Snelling RR, Fisher BL, Ward PS (eds) Advances in ant systematics (Hymenoptera: Formicidae): homage to E. O. Wilson—50 years of contributions. Plazi.orgGoogle Scholar
  25. Tschinkel WR (2006) The fire ants. The Belknap Press of Harvard University Press, CambridgeGoogle Scholar
  26. Valles SM, Porter SD (2003) Identification of polygyne and monogyne fire ant colonies (Solenopsis invicta) by multiplex PCR of Gp-9 alleles. Insectes Soc 50:199–200CrossRefGoogle Scholar
  27. Valles SM, Strong CA, Callcott A-MA (2016) Development of a lateral flow immunoassay for rapid field detection of the red imported fire ant, Solenopsis invicta (Hymenoptera: Formicidae). Anal Bioanal Chem 408:4693–4703CrossRefPubMedGoogle Scholar
  28. Valles SM, Wylie R, Burwell CJ, McNaught MK, Horlock C (2017) Evaluation of a lateral flow immunoassay for field identification of Solenopsis invicta (Hymenoptera: Formicidae) in Australia. Aust Entomol. CrossRefGoogle Scholar

Copyright information

© This is a U.S. government work and its text is not subject to copyright protection in the United States; however, its text may be subject to foreign copyright protection 2018

Authors and Affiliations

  • S. M. Valles
    • 1
    Email author
  • C. A. Strong
    • 1
  • A.-M. A. Callcott
    • 2
  1. 1.Center for Medical, Agricultural and Veterinary EntomologyUnited States Department of Agriculture, Agricultural Research ServiceGainesvilleUSA
  2. 2.Animal and Plant Health Inspection ServiceUnited States Department of Agriculture, Plant Protection and QuarantineBiloxiUSA

Personalised recommendations