Development of a molecular assay specific for the Leishmania donovani complex that discriminates L. donovani/Leishmania infantum zymodemes: a useful tool for typing MON-1.
We have developed a simple, rapid, sensitive, and cost-effective typing method, based on the amplicon size of the K26 gene, capable of species/strain discrimination of Leishmania donovani complex strains causing visceral leishmaniasis (VL). It was evaluated on 112 strains and compared with multilocus enzyme electrophoresis (MLEE) typing. The K26 polymerase chain reaction (PCR) applied on 26 representative L. donovani complex strains gave 14 different amplicon sizes. The assay was specific to the L. donovani complex and discriminated L. infantum from L. donovani strains. MON-1 strains were also easily distinguished from other non-MON-1. Surprisingly, 29.3% of the Greek strains included in this study were MLEE typed as MON-98 and gave exclusively a 940-bp amplicon. The majority of Greek MON-1 strains gave also the 940-bp amplicon, whereas a 626-bp amplicon was consistently obtained with other European MON-1 strains. K26 PCR-restriction fragment length polymorphism, based on MON-1 K26 sequence polymorphism, gave 2 MON-1 subgroups. Application of the method may contribute to efficiently monitor VL.
Haralambous
Christos
C
Laboratory of Molecular Parasitology, Department of Microbiology, Hellenic Pasteur Institute, 115 21 Athens, Greece.
Antoniou
Maria
M
Pratlong
Francine
F
Dedet
Jean-Pierre
JP
Soteriadou
Ketty
K
D000818:Animals; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004285:Dogs; D004592:Electrophoresis, Starch Gel; D004798:Enzymes / Q000032:analysis; D005838:Genotype; D006801:Humans; D007893:Leishmania donovani / Q000145:classification* / Q000235:genetics*; D018314:Leishmania infantum / Q000235:genetics*; D017720:Molecular Epidemiology / Q000191:economics / Q000379:methods*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015800:Protozoan Proteins / Q000235:genetics*; D017422:Sequence Analysis, DNA
D016054:DNA, Protozoan; D004798:Enzymes; D015800:Protozoan Proteins