Diagnostic enzymes are important for biochemical analysis of samples of blood, serum and urine for monitoring and diagnosing disease. With rising healthcare costs and to improve patient care, diagnostic laboratories have been challenged to develop new tests that are reliable, cost–effective and accurate and to optimize existing protocols by making them faster and more economical. Major step involves the immobilization of enzymes for their better handling and re-usability for screening large number of samples. In the present work, new method for fabrication of Polyvinyl chloride (PVC) based enzyme strips were developed that can be commercially exploited. PVC being cheaper, chemically inert with ease in moulding to various shapes & size will channel a new pathway in design & fabrication of diagnostic kits and related biomedical instrumentation.