Leishmania spp. parasite isolation through inoculation of patient biopsy macerates in interferon gamma knockout mice.
Isolation of Leishmania parasite and species identification are important for confirmation and to help define the epidemiology of the leishmaniasis. Mice are often used to isolate pathogens, but the most common mouse strains are resistant to infection with parasites from the Leishmania (Viannia) subgenus. In this study we tested the inoculation of interferon gamma knockout (IFNgamma KO) mice with biopsy macerates from Leishmania-infected patients to increase the possibility of isolating parasites. Biopsies from twenty five patients with clinical signs of leishmaniasis were taken and tested for the presence of parasites. Immunohistochemical assay (IHC) and conventional histopathology detected the parasite in 88% and 83% of the patients, respectively. Leishmania sp. were isolated in biopsy macerates from 52% of the patients by culture in Grace's insect medium, but 13% of isolates were lost due to contamination. Inoculation of macerates in IFNgamma KO mice provides isolation of parasites in 31.8% of the biopsies. Most isolates belong to L. (Viannia) subgenus, as confirmed by PCR, except one that belongs to L. (Leishmania) subgenus. Our preliminary results support the use of IFNgamma KO mice to improve the possibility to isolate New World Leishmania species.
Oliveira
Milton Adriano Pelli de
MA
Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goiás, Goiânia, Goiás, Brasil, 74605-050. mapoliv@usp.br
Pires
Alause da Silva
Ada S
de Bastos
Rosidete Pereira
RP
Lima
Glória Maria Collet de Araujo
GM
Pinto
Sebastião Alves
SA
Pereira
Ledice Inácia de Araujo
LI
Pereira
Ana Joaquina Cohen Serique
AJ
Abrahamsohn
Ises de Almeida
Ide A
Dorta
Miriam Leandro
ML
Ribeiro-Dias
Fátima
F
D000818:Animals; D001706:Biopsy; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics / Q000276:immunology; D007891:Leishmania / Q000145:classification / Q000302:isolation & purification*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout / Q000469:parasitology*; D016133:Polymerase Chain Reaction; D012867:Skin / Q000469:parasitology*; D013997:Time Factors
D007371:Interferon-gamma