Abstract Among all the modern technological advances, digital microfluidic biochip has been extending a salient solution to healthcare and bio-laboratories with the pledge of high sensitivity and reconfigurability. Such biochip devices fulfill the requirement of a faster testing kit for the detection of different novel diseases, which is indispensable in the market due to the tremendously disrupted scenario of healthcare systems. To eliminate erroneous testing, the current scope of digital microfluidic biochips is widened as a viable testing method using various bioprotocols with a reduced cost in developing countries. This paper addresses the existing security challenges and operational faults in the identification mechanism of proteins such as severe acute respiratory syndrome coronavirus 2 spike protein in state-of-the-art digital microfluidic biochips. We are the first to propose a safety detection solution along with a fault identification algorithm using an inductive transfer learning model. Experimental results of the proposed model register a threshold accuracy of 98% while applying the own dataset. This work will provide a better security-enabled fault-free safety assurance framework against attack and fault identification with better accuracy in digital microfluidic biochips for the detection of different diseases and many other healthcare diagnoses, without any overhead of completion time for bioprotocols.