PCR-Restriction Fragment Length Polymorfism Analysis (PRA) of Mycobacterium leprae

ZUMÁRRAGA, MARTÍN JOSÉ; RESOAGLI, E.H.; CICUTA, MARÍA ELENA; MARTÍNEZ A. R.; ORTIZ DE ROTT, M.I.; DE MILLÁN, S.G.; CAIMI, KARINA; GIOFFRÉ, ANDREA KARINA; ALITO, ALICIA; BIGI, FABIANA; CATALDI, ANGEL; ROMANO, MARÍA ISABEL

INTERNATIONAL JOURNAL OF LEPROSY AND OTHER MYCOBACTERIAL DISEASES

International Leprosy Association

Año: 2001 vol. 69 p. 21 - 25

0148-916X

Polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis (PRA) which relies on the amplification of a 439-bp portion of the hsp65 gene present in all mycobacteria, followed by two distinct digestions (with BstEII and HaeIII) of the PCR product, offers a rapid and easy alternative that allows identification of the species without the need for specialized equipment. Wild leprosy in the nine-banded armadillo (Dasypus novemcinctus) is characterized by the presence of multiple bacilli in internal organs such as lymph nodes, spleen and liver, as well as in nerves and skin. We could observe this in 9 out of 132 animals captured in Corrientes, Argentina, an area endemic for leprosy in humans. Mycobacterium leprae were recognized in those naturally infected animals through different techniques. Three samples of extracted DNA of the mycobacteria present in the spleen, liver and popliteal lymph node of a naturally infected animal during the Experimental Program in Armadillo (PEA) and three samples of human lepromas were processed by PRA. The patterns of the six samples analyzed were identical and were characteristic of M. leprae. These studies, made for the first time in Argentina, corroborate the initial discoveries in South America made by our investigative group on the detection of armadillos naturally infected with the Hansen bacillus.