Leishmania identification by PCR of Giemsa-stained lesion imprint slides stored for up to 36 years.
This study examined the ability of PCR to amplify Leishmania DNA, stored on Giemsa-stained slides, from American cutaneous leishmaniasis (ACL) patients. In total, 475 slides stored for up to 36 years were obtained from an outpatient clinic in a Brazilian ACL-endemic region, and Leishmania DNA was amplified from 395 (83.2%) of the DNA samples using primers specific for the minicircle kinetoplast DNA. Restriction fragment length polymorphism analysis of these amplicons demonstrated that Leishmania (Viannia) braziliensis was the only species present in these samples. The results demonstrated that archived Giemsa-stained slides can provide a Leishmania DNA source for performing clinical and epidemiological studies of leishmaniasis.
Volpini
A C
AC
Laboratório de Pesquisas em Leishmanioses, Departamento de Imunologia, IOC-FIOCRUZ, Rio de Janeiro, Brazil. avolpini@ioc.fiocruz.br
Marques
M J
MJ
Lopes dos Santos
S
S
Machado-Coelho
G L
GL
Mayrink
W
W
Romanha
A J
AJ
D000818:Animals; D001399:Azure Stains; D016054:DNA, Protozoan / Q000032:analysis; D006801:Humans; D007892:Leishmania braziliensis / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D013048:Specimen Handling; D013997:Time Factors
D001399:Azure Stains; D016054:DNA, Protozoan