Phylogenic analysis of the genus Leishmania by cytochrome b gene sequencing.
In a previous report (Luyo-Acero et al., 2004), we demonstrated that cytochrome b (Cyt b) gene analysis is an effective method for classifying several isolates of the genus Leishmania; hence, we have further applied this method to other Leishmania species in an effort to enhance the accuracy of the procedure and to construct a new phylogenic tree. In this study, a total of 30 Leishmania and Endotrypanum WHO reference strains, clinical isolates from our patients assigned to 28 strains (human and non-human pathogenic species) and two species of the genus Endotrypanum were analyzed. The Cyt b gene in each sample was amplified by PCR, and was then sequenced by several primers, as reported previously. The phylogenic tree was constructed based on the results obtained by the computer software MEGA v3.1 and PAUP* v4.0 Beta. The present phylogenic tree was almost identical to the traditional method of classification proposed by Lainson and Shaw (1987). However, it produces the following suggestions: (1) exclusion of L. (Leishmania) major from the L. (L.) tropica complex; (2) placement of L.tarentolae in the genus Sauroleishmania; (3) L. (L.) hertigi complex and L. (V.) equatorensis close to the genus Endotrypanum; (4) L. (L.) enrietti, defined as L. (L.) mexicana complex, placed in another position; and (5) L. (L.) turanica and L. (L.) arabica are located in an area far from human pathogenic Leishmania strains. Cyt b gene analysis is thus applicable to the analyzing phylogeny of the genus Leishmania and may be useful for separating non-human pathogenic species from human pathogenic species.
Asato
Yutaka
Y
Division of Dermatology, Unit of Organ-oriented Medicine, University of the Ryukyu, 207 Uehara, Nishihara-cho, Okinawa 903-0215, Japan. yasatoy@med.u-ryukyu.ac.jp
Oshiro
Minoru
M
Myint
Chomar Kaung
CK
Yamamoto
Yu-ichi
Y
Kato
Hirotomo
H
Marco
Jorge Diego
JD
Mimori
Tatsuyuki
T
Gomez
Eduardo A L
EA
Hashiguchi
Yoshihisa
Y
Uezato
Hiroshi
H
D000818:Animals; D001483:Base Sequence; D016384:Consensus Sequence; D045303:Cytochromes b / Q000235:genetics*; D016054:DNA, Protozoan / Q000737:chemistry*; D006801:Humans; D007891:Leishmania / Q000145:classification* / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D014351:Trypanosomatina / Q000145:classification / Q000235:genetics
D016054:DNA, Protozoan; D045303:Cytochromes b