Consistence of miniexon polymerase chain reaction-restriction fragment length polymorphism and single-copy gene sequence analyses in discriminating Leishmania genotypes.
Recently, a new polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP)-based assay had been developed using the miniexon sequences for genotyping Leishmania isolates. We had used this method for rapid diagnosis and genotyping of visceral and cutaneous leishmaniasis with the combination of microcapillary cultivation. In this study, we have evaluated this approach by examining genomic DNAs from 47 independent isolates, which were grouped into 19 genotypes of Leishmania subgenus complexes by sequence polymorphism of single-copy genes. Results obtained provide miniexon RFLP configurations specific to Leishmania enriettii, Leishmania tarentolae, and Leishmania gerbilli for the first time. Altogether, 92% of the results from miniexon PCR-RFLP are in agreement with those based on the sequence database of single-copy genes from the same isolates. The miniexon PCR-RFLP method is simple, sensitive, and specific method useful for routine diagnosis of different Leishmania.
Serin
Mehmet S
MS
Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Mersin University, 33343 Yenisehir, Mersin, Turkey. serinm@mersin.edu.tr
Waki
Kayoko
K
Chang
Kwang-Poo
KP
Aslan
Gonul
G
Direkel
Sahin
S
Otag
Feza
F
Kayar
Begum
B
Koksal
Fatih
F
Emekdas
Gurol
G
D000818:Animals; D016054:DNA, Protozoan / Q000032:analysis; D005091:Exons / Q000235:genetics*; D005838:Genotype; D006801:Humans; D007891:Leishmania / Q000145:classification* / Q000235:genetics*; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic / Q000235:genetics; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity
D016054:DNA, Protozoan