Heat Treatment Augments Antigen Detection of Dirofilaria immitis in Apparently Healthy Companion Dogs (3.8% to 7.3%): Insights from a Large-Scale Nationwide Survey across the United States

Background: Heartworm disease (HWD) is a vector-borne disease caused by the filarial nematode Dirofilaria immitis. Low antigen levels caused by immune complex formation preclude HWD diagnosis. Heat treatment is an immune complex dissociation technique used to enhance antigen detection. Only a few studies have reported the benefits of heat treatment in nationwide surveys. Methods: To investigate the impact of heat treatment on the seroprevalence of HWD in companion dogs in the USA, serum samples (n = 3253) were analyzed for D. immitis antigen (DiroCHEK (R), Zoetis) without and with heat treatment of the samples. Results: Compared to sera without heat treatment, heat treatment significantly increased overall prevalence from 3.8% (123/3253) to 7.3% (237/3253) (p < 10(-4)), expanding antigen detection from 32 to 39 of the 48 states and Washington District of Columbia included in this study. Conclusions: This study represents the largest nationwide survey of HW antigen detection in dogs in the US applying heat treatment to canine sera. The heat treatment used herein has the advantage of requiring a low volume of serum, making it optimal for use in routine diagnosis. Heat treatment should be used routinely by reference laboratories and veterinary clinics in patients with a negative initial test.