The vets refused the request from a property owner in Landsborough, forcing the state's Department of Primary Industries to send its own vet.
Acting chief veterinary officer Dr Rick Symons said tests on the dead horse ultimately proved negative for the virus.
"Based on information from the owner and the veterinarian involved with the property, the horse was considered low risk for Hendra virus and the test results confirm this," he said.
"This case was unique in that the owner asked two local vets to take samples from the dead horse."
After they declined, an experienced Biosecurity Queensland veterinarian visited the property on Monday night to take the samples.
Dr Symonds said Biosecurity Queensland was working with the Australian Veterinary Association and workplace health and safety authorities to educate vets in infection control and the use of personal protective equipment to ensure they were confident and prepared to deal with Hendra-suspect horses.
"Biosecurity Queensland will respond to suspect cases deemed to be high risk and positive cases of Hendra virus. It is not normal practice for the agency to collect samples from horses that are assessed as low risk for Hendra virus," he said.
"Biosecurity Queensland receives up to 10 samples each day from private veterinarians across Queensland and it would be impractical for government veterinarians to attend each case to collect samples.
"The Biosecurity Queensland laboratory processes samples submitted from private veterinarians from such cases.
"Biosecurity Queensland continues to take any suspect cases of Hendra virus cases extremely seriously and will assist where possible."