title: Rapid diagnosis and genotyping of Leishmania isolates from cutaneous and visceral leishmaniasis by microcapillary cultivation and polymerase chain reaction-restriction fragment length polymorphism of miniexon region.
abstract: We have performed a combination of microcapillary cultivation method and restriction fragment length polymorphism (RFLP) analysis of amplified products by 1 single PCR of miniexon region of Leishmania for molecular diagnosis and genotyping of different Leishmania species isolated from cutaneous leishmaniasis (CL) and visceral leishmaniasis. We have analyzed 10 microcapillary cultivated isolates from cutaneous cases and 5 microcapillary cultivated isolates from visceral cases (totally 15) by polymerase chain reaction-RFLP (PCR-RFLP). Of 10 isolates, 3 (30%) were genotyped as Leishmania infantum and 7 (70%) of 10 isolates were genotyped as Leishmania tropica from the microcapillary cultivated isolates of cutaneous cases. On the other hand, all 5 isolates (100%) were genotyped as L. infantum from microcapillary cultivated visceral cases. Our most interesting finding is the presence of 3 L. infantum isolates in CL cases without kala-azar history. Therefore, we suggest that further investigations must be done about this subject. On the other hand, we suggest the combination of microcapillary culture method and PCR-RFLP of miniexon region of leishmaniae can be used in routine laboratory experimentation because of their simple, cheap, and rapid benefits (within a week), whereas other different approaches offer a multitude of valid taxonomic characters for species identification.
authors.affiliation: Department of Microbiology, Faculty of Pharmacy, Mersin University, 33343, Turkey. serinmss@cu.edu.tr
authors.forename: Mehmet S
authors.initials: MS
authors.lastname: Serin
authors.forename: Kenan
authors.initials: K
authors.lastname: Daglioglu
authors.forename: Melahat
authors.initials: M
authors.lastname: Bagirova
authors.forename: Adil
authors.initials: A
authors.lastname: Allahverdiyev
authors.forename: Soner
authors.initials: S
authors.lastname: Uzun
authors.forename: Zeynep
authors.initials: Z
authors.lastname: Vural
authors.forename: Begum
authors.initials: B
authors.lastname: Kayar
authors.forename: Seda
authors.initials: S
authors.lastname: Tezcan
authors.forename: Mesut
authors.initials: M
authors.lastname: Yetkin
authors.forename: Gonul
authors.initials: G
authors.lastname: Aslan
authors.forename: Gurol
authors.initials: G
authors.lastname: Emekdas
authors.forename: Fatih
authors.initials: F
authors.lastname: Koksal
mesh_terms: D000818:Animals; D003470:Culture Media; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005091:Exons / Q000235:genetics; D006801:Humans; D007891:Leishmania / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000469:parasitology; D007898:Leishmaniasis, Visceral / Q000175:diagnosis* / Q000469:parasitology; D010274:Parasitology / Q000295:instrumentation / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D013997:Time Factors
chemical_list: D003470:Culture Media; D015252:Deoxyribonucleases, Type II Site-Specific; C037794:YGGCCR-specific type II deoxyribonucleases