Produção de polipeptídeo recombinante para utilizacão no diagnóstico sorológico da hanseníase

Abstract

Leprosy also known as leprosy or Hansen's disease is caused by the alcohol-acid bacillus Mycobacterium leprae, is endemic in several parts of the world, affecting thousands of people in several countries. There is an operational classification established by WHO for treatment purposes, in which the disease is divided into paucibacillary and multibacillary. Treatment based on long-term polychemotherapy is stipulated according to this classification. The diagnosis for leprosy is based on clinical evaluation, since existing tests are unreliable and usually unable to detect individuals from the paucibacillary pole of the disease. A reliable diagnostic method is essential for a better control and reduction of the epidemiological indices of the disease. The aim of this work was to produce and purify a recombinant M. leprae polypeptide that demonstrates in vitro leprosy detection ability. The gene encoding a synthetic polypeptide (MLP-15) containing 15 peptides from M. leprae was designed using Vector-NTI software and synthesized by the company Epoch Biolab Company®. In silico analyzes evaluating the physico-chemical parameters, secondary and tertiary structure, besides the capacity of detection by MHC of classes I and II were performed. MLP-15 was cloned and expressed in E. coli using BL21 strain RP and purified with Ni-NTA affinity column in sufficient quantity to be used in diagnostic tests. All parameters evaluated in silica indicated that MLP-15 has potential to be used in serological tests for the detection of leprosy patients.