A Preliminary Study Aimed at the Detection of Leishmania Parasites in Subjects with Cutaneous Leishmaniasis Using Polymerase Chain Reaction

Abstract

As a basic study for future diagnosis of cutaneous leishmaniasis, we tried to detect Leishmania parasites representing different species in the subgenera Leishmania and Viannia from subject patients with cutaneous leishmaniasis by using the polymerase chain reaction (PCR) with the subgenus Viannia specific primer. Four out of the 14 specimens revealed an amplified DNA of 70 bp specific for the subgenus Viannia (L. braziliensis complex). No bands were detected in the rest of the specimens belonging to the subgenus Leishmania and unclassified groups. The base sequences of the amplified DNA corresponded with those of the L. (V). braziliensis kinetoplast minicircle. We concluded that PCR using the present primer specific for the subgenus Viannia would be useful in detecting Leishmania parasites in lesions of cutaneous leishmaniasis caused by the L. braziliensis complex.

Low-Dose UVB Contributes to Host Resistance againstLeishmania amazonensisInfection in Mice through Induction of Gamma Interferon and Tumor Necrosis Factor Alpha Cytokines

“…A total of 20 mice in each group were used. L. amazonensis (MHOM/ BR/73/M2269) parasites were cultured in vitro as reported previously (45).…”

“…In contrast, at 6 h after the last irradiation and in control mice, negative results were seen for TNF-␣. The same method of PCR repeatedly failed to yield a signal for IL-4, despite an increase in the number of cycles to 45.…”

Diagnostic approach and significance of inducible nitric oxide positivity in human cutaneous leishmaniasis caused by leishmania tropica

“…WHO has designated leishmaniasis as one of six major diseases, with the possibility of becoming more widespread in the near future 27 . The current treatment of leishmaniasis is expensive and toxic.…”

Detection of Species of the Subgenus Leishmania Parasites Using Polymerase Chain Reaction and Southern Blotting

“…They were cultured in vitro in RPMI 1640 (Nakaraitesuqu, Japan) medium containing 10% heat inactivated fetal bovine serum (FBS) (Bio Whittaker, USA) supplemented with antibiotics (30 μg/ml Ampicillin+100 μg/ml Gentamycin) at 25°C. Genomic DNA was extracted from cultured promastigotes as described previously (12, 13).…”

“…A number of PCR based methods for diagnosis of leishmaniases by detecting parasite genomic DNA have been reported with varying specificity (9‐11); but only a few could identify the parasite precisely. In our previous studies, we developed a method for species identification of subgenus Viannia from clinical samples using PCR and Southern blotting analysis (12, 13). In the present report, we have extended these previous studies and attempted to identify the Leishmania species of the subgenus Leishmania precisely by applying PCR and Southern blotting.…”