Improved Antigen Detection of Male-Only Dirofilaria immitis Infections in Canine Serum after Heat Treatment for Immune Complex Dissociation

Abstract

Since the mid-1990s, male-only heartworm infections have been considered undetectable using antigen tests based on experimental studies. Results from those studies are in contrast to reports in the decade prior showing variable male heartworm antigen detection using naturally infected animals and antigen tests using chemical and/or heat immune complex dissociating steps. Several recent studies utilizing heat treatment for immune complex dissociation (Heat ICD) demonstrated increased antigen sensitivity for necropsy verified male-only infections and a higher-than-expected frequency of this type of infection. This study utilized archived canine serum with verified male-only heartworm infections to evaluate detection of the heartworm antigen using the DiroCHEK® (Zoetis LLC, Parsippany, NJ, USA), Witness® (Zoetis LLC, Parsippany, NJ, USA), and SNAP® Heartworm RT (IDEXX Laboratories, Inc., Westbrook, ME, USA) antigen tests. Results showed significant increases in sensitivity for the heartworm antigen following heat treatment for DiroCHEK® (+42.1%, p < 0.0001) and Witness® (+26.3%, p = 0.0020), but not the SNAP® Heartworm RT (+10.5%, p = 0.1250). Prior to heat treatment, false negative results were obtained in 76.3–83.0% of mature infections. Infections with only immature male worms were never detected using any heartworm test used. Heat treatment of serum allows improved detection of mature male-only heartworm infections, which may occur more frequently than previously recognized, and like all heartworm infections pose a risk of chronic and cumulative pathology as well as thromboembolic disease regardless of infection intensity.