Comparative Evaluation of 2 In-Clinic Assays for Vector-Borne Disease Testing in Dogs

Abstract Vector-borne agents comprise medically important infections affecting dogs throughout much of the world. Sensitive detection of antibodies directed at tick-borne disease-causing organisms in dogs is diagnostically important for veterinarians, pets and their owners, and epidemiologically important for public health surveillance. The SNAP 4Dx Plus Test (IDEXX Laboratories, Inc., Westbrook, ME) identifies antibodies to or infection with multiple tick-borne pathogens and canine heartworm antigen in a single assay. Recently, VetScan FLEX4 Rapid Test (Abaxis, Inc., Union City, CA) was launched as a new assay to detect tick-borne pathogen antibodies and heartworm antigen. In the present study, we evaluated the comparative performance of SNAP 4Dx Plus (SNAP) and FLEX4 Rapid Test (FLEX4) using samples selected based on geographic distributions for canine vector borne diseases, including Borrelia burgdorferi ( n = 105), Anaplasma phagocytophilum (160), Anaplasma platys (115), Ehrlichia canis (154), Ehrlichia ewingii (163), Ehrlichia chaffeensis (151) and Dirofilaria immitis (105). Canine vector borne diseases infection status was established for each sample by a combination of reference methods that included necropsy ( D. immitis, heartworm disease), Western immunoblotting ( B. burgdorferi ), immunofluorescence assays ( A. phagocytophilum and E. canis ) and species-specific ELISAs ( A. platys, E. canis, E. ewingii and E. chaffeensis ). For comparisons among the 2 assays, samples were evaluated per the manufacturers’ instructions for each test kit. By testing each same sample set compared to the defined reference results, sensitivities differed substantially between SNAP and FLEX4, at 95.5 vs. 40.9%, respectively for B. burgdorferi , 97.1% vs. 61.4% for E. canis , 98.2% vs. 59.3% for E. ewingii , 64.3% vs. 35.7% for E. chaffeensis , 84.5% vs. 12.7% for A. phagocytophilum , 83.3% vs. 33.3% for A. platys , and 94.1% vs. 88.2% for D. immitis . Specificities for both rapid assay tests ranged from 98% to 100%. Based upon the comparative results derived from this study, the SNAP test was more sensitive than the FLEX4 test for detection of antibodies to all tick-borne pathogens and heartworm disease ( Dirofilaria immitis ) antigen in dogs.