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Background  Detection of  Dirofilaria immitis , or heartworm, through antigen in sera is the primary means of diagnosing infections in dogs. In recent years, the practice of heat-treating serum prior to antigen testing has demonstrated improved detection of heartworm infection. While the practice of heat-treating serum has resulted in earlier detection and improved sensitivity for heartworm infections, it has been suggested that heat treatment may cause cross reactivity with  A. reconditum  and intestinal helminth infections of dogs. No studies have assessed the potential cross-reactivity of these parasites with heartworm tests before and after heat treatment using blood products and an appropriate gold standard reference.    Methods  Canine sera ( n =163) was used to evaluate a heartworm antigen-ELISA (DiroCHEK®) and potential cross-reactivity with common parasitic infections. The heartworm status and additional parasite infections were confirmed by necropsy and adult helminth species verified morphologically or by PCR, and feces evaluated by centrifugal fecal flotation.    Results  Intestinal parasites were confirmed in 140 of the dogs by necropsy, and 130 by fecal flotation.  Acanthocheilonema reconditum  microfilariae were confirmed in 22 dogs. Prevalence of heartworm infection confirmed by necropsy was 35.6% (58/163). In the 105 dogs without heartworms, specificity remained unchanged at 100% both before and after heat treatment despite confirmed infections with  A. reconditum ,  Ancylostoma caninum ,  Ancylostoma brasiliense ,  Trichuris vulpis ,  Toxocara canis ,  Dipylidium caninum ,  Spirometra mansonoides ,  Macracanthorynchus ingens ,  Cystoisospora  sp.,  Giardia  sp., and  Sarcocystis  sp.    Conclusions  These findings suggest that the use of heat treatment improves sensitivity of heartworm tests and is unlikely to cause false positive antigen results due to  Acanthocheilonema reconditum , intestinal helminths, and protozoal parasites in dogs.Graphic abstract

Detection of heartworm antigen without cross-reactivity to helminths and protozoa following heat treatment of canine serum