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

Abstract

In this study, an attempt was made to identify different Leishmania species by polymerase chain reaction (PCR). Fourteen Leishmania strains from stock were tested by PCR and Southern blotting. A pair of primers were employed that anneal to the kinetoplast DNA sequence conserved among subgenus Leishmania. Of the 14 Leishmania strains used in this study, six showed strong bands of approximately 170 bp, and all the positive strains belonged to the species of the subgenus Leishmania viz., Leishmania (Leishmania) garnhami, L. (L.) amazonensis, L. (L.) pifanoi, L. (L.) mexicana, L. (L.) chagasi, and L. (L.) major All the species belonging to the subgenus Viannia used in this study were negative by PCR. These results suggest that the primer pair may be useful for identification of the species belonging to the subgenus Leishmania of the New World as well as to distinguish subgenus Leishmania from subgenus Viannia.

Single step polymerase chain reaction (PCR) for the diagnosis of the Leishmania (Viannia) subgenus

“…Several authors reported molecular approaches for identifying the subgenus and species of Leishmania (CASTILHO et al, 2003;RODRIGUES et al, 2002;UEZAT et al, 2001;FERNANDES et al, 1999) but some of these methods are either laborious or the DNA sequences used as target are usually present as single or low copy. Based on these reports, a comparison was undertaken between LV1 and LV2 mini-exon PCR and PCR-RFLP (VOLPINI et al, 2004).…”

An imported case of cutaneous leishmaniasis caused by Leishmania (Leishmania) donovani in Japan

“…Leishmaniasis is an infection caused by Leishmania protozoa and is transmitted by the bite of a sandfly . There are 21 different Leishmania species, and identification of the causative Leishmania species is very important, because clinical symptoms, prognosis and response to medication are different depending on the infecting Leishmania species . Leishmaniasis is classified into three clinical forms: cutaneous leishmaniasis (CL), mucocutaneous leishmaniasis (MCL) and visceral leishmaniasis (VL) .…”