A questionnaire was developed to document the knowledge base of large-animal diplomates of the American College of Veterinary Internal Medicine (ACVIM) regarding polymerase chain reaction (PCR) technology and to identify the common use of this technology in equine practice. Ninety-three of the 278 mailed questionnaires were returned, for an overall response rate of 33.4%. Ninety respondents (99%) reported being familiar with the general principles of nucleic acid probe technology; however, only 52 (57%) knew the difference between conventional (traditional) and real-time (second-generation) PCR. The majority of the respondents (88%) emphasized the need for continuing education on molecular diagnostics. Eighty-four (92%) of the respondents regularly use PCR (conventional and/or real-time) for the detection of equine pathogens, and 80 (88%) commonly submit their samples to university/state veterinary laboratories. Blood, nasal swabs, and feces are the three equine specimens most commonly submitted for PCR analysis of Streptococcus equi, Lawsonia intracellularis, Neorickettsia risticii. equine herpesvirus 1/4, Rhodococcus equi, Sarcocystis neurona, and equine influenza virus. Diplomates reported costs associated with molecular diagnostics and unreliability of PCR as the most common limitations of PCR. Didactic training in veterinary curricula and during continuing-education opportunities continues to be necessary to produce veterinarians who have an understanding of the clinical applications of molecular diagnostics.
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