The development of probe based qPCR assay for CYP39A1 G204E mutation detection

Author: Bachana Kapanadze
Keywords: Exfoliation Syndrome, TaqMan probe, polymerase chain reaction, mutation.
Annotation:

Exfoliation syndrome is a systemic age-related disorder and the most common cause of glaucoma and irreversible blindness. As it turns out, individuals with exfoliation syndrome have twice the risk of having certain mutations in the gene encoding the CYP39A1 protein, an enzyme which has an important role in the metabolism of cholesterol. The risks of developing exfoliation syndrome are associated with the CYP39A1 G204E mutation, which notably increases the severity of the disease. Patients with exfoliation syndrome carrying the CYP39A1 G204E mutation have a significantly increased risk of developing glaucoma and irreversible blindness compared to patients with exfoliation syndrome without any mutation. The characteristics of exfoliation syndrome, its treatment methods and diagnostic assays are especially current in the modern world because, as of 2020, glaucoma was recorded in about 80 million people worldwide. According to statistical data, this number will probably increase up to 111 million by 2040. Currently, the characteristics and exact mechanisms of the pathogenesis of exfoliation syndrome are not fully understood and remains as the subject of active research in ophthalmology. Based on the relevance of exfoliation syndrome, the project aimed to design and develop a probe-based polymerase chain reaction (qPCR) assay and protocol for the detection of CYP39A1 G204E mutation in order to evaluate the risks of development of irreversible blindness and glaucoma in patients with exfoliation syndrome carrying a mutation in the CYP39A1 gene and without a mutation in the CYP39A1 gene. During the development of the diagnostic assay, selected target object was human nuclear DNA. Time and cost efficient probe based polymerase chain reaction will definitely help patients detect the mutation at an early stage in case of existing that will open up the opportunity to take appropriate measures in order to alleviate the course of disease. The diagnostic assay developed within the framework of our project will open up new perspectives in the diagnosis of exfoliation syndrome.