Development of a Screening Kit for Early Diagnosis and Prevention of Wilson´s Disease

Abstract

Purpose : Wilson´s disease is an autosomal recessive disorder characterized by copper accumulation in the liver, brain, and other organs due to defected copper metabolism. The incidence of Wilson´s disease is approximately one in 30,000 population in the world, more common than phenylketonuria in Korea. The early diagnosis or presymptomatic diagnosis of Wilson´s disease is critical in order for them to live a normal life. However, unfortunately, there are no commercial kits available for Wilson´s disease screening in the world yet. Methods : We developed a mass-screening kit for the purpose of early diagnosis and prevention of Wilson´s disease using sandwich ELISA method. This kit can handle a large number of samples at the same time by using filter paper as in newborn screening. Using the polyclonal or monoclonal anti-ceruloplasmin antibodies, this kit determines the plasma ceruloplasmin levels-one of the main markers for Wilson´s disease. Results : The plasma levels of the ceruloplasmin were considerably lower in the Wilson´s disease (4.5±1.6 mg/dL) group compared to normal controls(22.1±1.4 mg/dL), sufficient to be used for mass screening. In addition, the results using this screening kit showed 100% positive and negative concordance rates with the test results obtained from immuno-turbidimetry analysis which is the currently used in most test centers for ceruloplasmin measurement in the serum or plasma after centrifugation. Conclusion : Taken together, we successfully developed a screening kit which is very effective for the early diagnosis and prevention of Wilson´s disease. By using simple filter paper method for sample collection, this kit provides suitable mass screening. We suggest the screening for Wilson´s disease at the age of 3-5 years.