New primers for the detection Leishmania species by multiplex polymerase chain reaction

• Published in: Parasitology research (1987) Vol. 117; no. 2; pp. 501 - 511
• Main Authors: Conter, Carolina Cella, Lonardoni, Maria Valdrinez Campana, Aristides, Sandra Mara Alessi, Cardoso, Rosilene Fressatti, Silveira, Thaís Gomes Verzignassi
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-02-2018; Springer; Springer Nature B.V
• Subjects: Biomedical and Life Sciences; Biomedicine; Decision making; Geographical distribution; Health aspects; Immunology; Leishmania; Leishmaniasis; Medical Microbiology; Microbiology; Observations; Original Paper; Parasites; Parasitic diseases; Polymerase chain reaction; Primers; Protozoa; Scientific apparatus & instruments; Species; Spleen; Polymerase chain reaction; Multiplex PCR; species; Primers; DNA; Diagnostic; Leishmaniasis; Leishmania species
• ISSN: 0932-0113; 1432-1955
• DOI: 10.1007/s00436-017-5726-1

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.