Objective To develop a biosafety negative?pressure poultry isolator. Methods In accordance with national standards for animal isolation equipment, taking into account the actual needs of airtight transfer and disinfection functions, the structure of an isolator was designed. Results The isolator was integrated with an operating system, ventilation and filtration system, transfer system, automatic control system, sterilization connection system, and sewage collection and treatment system. Upon testing by a third party, it was confirmed that its technical indicators meet the national standards. Conclusions The isolator is suitable for feeding and experimental research of animals with highly pathogenic infections, meeting the practical needs of biosafety laboratories in China.