Are cytochrome B gene sequencing and polymorphism-specific polymerase chain reaction as reliable as multilocus enzyme electrophoresis for identifying Leishmania spp. from Argentina?
Recently, two techniques, polymerase chain reaction (PCR) amplification and sequencing of cytochrome b gene (cyt b gene sequencing) and polymorphism-specific PCR (PS-PCR) were recommended for Leishmania species identification. Before this study, however, the accuracy of these methods had not been tested against the multilocus enzyme electrophoresis, the current gold standard technique on this task. Therefore, a trial was done for the first time to compare the results obtained by these techniques, using 17 Argentinean Leishmania stocks in independent assays. For all the stocks examined, the same results at species level were obtained by the three techniques. Among them, 14 were assigned to L. (Viannia) braziliensis, and three to L. (V.) guyanensis. The two techniques, cyt b gene sequencing and PS-PCR, were able to distinguish between all the proven species responsible for leishmaniases in Argentina. Thus, both techniques were validated and could be used independently for the species designation of Leishmania parasites in the country.
Marco
Jorge D
JD
Department of Parasitology, Kochi Medical School, Kochi University, Kohasu, Kochi, Japan. marcojd@med.kochi-ms.ac.jp
Uezato
Hiroshi
H
Mimori
Tatsuyuki
T
Barroso
Paola A
PA
Korenaga
Masataka
M
Nonaka
Shigeo
S
Basombrío
Miguel A
MA
Taranto
Néstor J
NJ
Hashiguchi
Yoshihisa
Y
D000818:Animals; D001118:Argentina; D001483:Base Sequence; D045303:Cytochromes b / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry; D004311:Double-Blind Method; D004587:Electrophoresis, Agar Gel; D007891:Leishmania / Q000145:classification* / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards*; D011110:Polymorphism, Genetic; D015203:Reproducibility of Results; D016415:Sequence Alignment / Q000379:methods; D013045:Species Specificity
D017931:DNA Primers; D045303:Cytochromes b