Identification and determination of the relationships of species and strains within the genus Leishmania using single primers in the polymerase chain reaction.
DNA polymorphisms were assessed in different species and strains within the genus Leishmania by amplifying genomic DNA with single non-specific primers. This polymerase chain reaction (PCR) method employed non-random primers which anneal to mini- and microsatellite DNA sequences like the M13 core sequence and the simple repeat sequences (GTG)5 and (GACA)4, and the T3B primer derived from an intergenic spacer for tRNA genes. Distinctive and reproducible sets of amplified DNA fragments were obtained for all Leishmania isolates tested. The number and size of amplification products were found to be characteristic for a given taxon. Highly similar PCR profiles were observed when genomic DNA of representatives of the L. donovani, L. mexicana or L. braziliensis complexes was amplified. By comparing PCR patterns of unidentified Leishmania isolates with those obtained from reference strains it was possible to identify these isolates at the species level. The information of the amplification patterns was used for the construction of phylogenetic trees to measure the genetic relatedness within the genus Leishmania.
Schönian
G
G
Institut für Mikrobiologie und Hygiene, Universitätsklinikum Charité, Humboldt-Universität zu Berlin, Germany. schoeni@rz.charite.hu-berlin.de
Schweynoch
C
C
Zlateva
K
K
Oskam
L
L
Kroon
N
N
Gräser
Y
Y
Presber
W
W
D000818:Animals; D017931:DNA Primers; D016054:DNA, Protozoan / Q000235:genetics; D005843:Geography; D007438:Introns; D007891:Leishmania / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D016053:RNA, Protozoan / Q000235:genetics; D012343:RNA, Transfer / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D018911:Trinucleotide Repeats
D017931:DNA Primers; D016054:DNA, Protozoan; D016053:RNA, Protozoan; D012343:RNA, Transfer