| Autor: |
Conter CC; Graduate Program in Health Sciences, State University of Maringá, 5790, Colombo Avenue, Maringá, Paraná, 87020-900, Brazil., Lonardoni MVC; Department of Clinical Analysis and Biomedicine, State University of Maringá, 5790, Colombo Avenue, Paraná, Maringá, 87020-900, Brazil., Aristides SMA; Department of Clinical Analysis and Biomedicine, State University of Maringá, 5790, Colombo Avenue, Paraná, Maringá, 87020-900, Brazil., Cardoso RF; Department of Clinical Analysis and Biomedicine, State University of Maringá, 5790, Colombo Avenue, Paraná, Maringá, 87020-900, Brazil., Silveira TGV; Department of Clinical Analysis and Biomedicine, State University of Maringá, 5790, Colombo Avenue, Paraná, Maringá, 87020-900, Brazil. tgvsilveira@uem.br. |
| Abstrakt: |
Leishmaniasis is caused by protozoa of the Leishmania genus, which is divided into subgenus Viannia and Leishmania. In humans, the course of infection largely depends on the host-parasite relationship and primarily of the infective species. The objective of the present study was to design specific primers to the identification of Leishmania species using multiplex PCR. Four primers were designed, based on the GenBank sequences of the kDNA minicircle, amplifying 127 bp for subgenus Viannia, 100 bp for L. amazonensis, and 60 bp for Leishmania donovani complex and L. major. None of the primers amplified Trypanosoma cruzi or L. mexicana. The limit of detection of multiplex PCR was 2 × 10 -5 parasites for L. braziliensis, 2 x 10 -3 parasites for L. amazonensis, and 1.4 × 10 -3 parasites for L. infantum. The high sensitivity of multiplex PCR was confirmed by the detection of parasites in different biological samples, including lesion scrapings, spleen imprinting of a hamster, sandflies, and blood. The multiplex PCR that was developed herein presented good performance with regard to detecting and identifying the parasite in different biological samples and may thus be useful for diagnosis, decision making with regard to the proper therapeutic approach, and determining the geographic distribution of Leishmania species. |
Full text from SpringerLink