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It has been documented that the genotypic traits in symptomatic and asymptomatic cases of visceral leishmaniasis (VL) may be different. The current study aimed to find out and compare the genotype and intraspecies diversity of  Leishmania  Internal Transcribed Spacer 1 (ITS1) from asymptomatic and symptomatic VL cases in southern Iran.  Methods . Buffy coat samples from seven VL patients, with clinical signs and symptoms, and seven asymptomatic VL cases, were evaluated in this study. Samples of asymptomatic individuals were obtained from children living in a VL endemic area in southern Iran, while the samples of symptomatic subjects were obtained from patients admitted to hospitals with a diagnosis of VL. DNA was extracted from the buffy coats of the samples and PCR-amplified, targeting the ITS1of  Leishmania . The PCR products were sequenced, and the consensus sequences were assembled and multiple-aligned with a set of  Leishmania  strains retrieved from the GenBank, using Clustal W. The phylogenetic tree was rooted, using MEGAX software, and the diversities based on haplotype and nucleotides, as well as the number of polymorphic sites, were measured using DnaSP v5.0 software. The results of ITS1 sequencing in 5 out of 7 asymptomatic VL cases showed 99.25% to 100% similarity with the  Leishmania infantum  ITS1 sequence (accessed number: MN648746), and one isolate was considered as just  Leishmania  sp. In one sample, 99.75% similarity was seen with the ITS1 sequence of  Crithidia fasciculata . Of the symptomatic VL patients, the PCR product revealed a 340 bp band corresponding to  L. infantum  in all of the samples. By analyzing the ITS1 sequences, all seven sequences formed a clade somewhat different from other  Leishmania  species and considered as  Leishmania  sp. Haplotype and nucleotide diversity were much more prevalent in symptomatic cases where six haplotypes were seen in the ITS1 of  Leishmania  from symptomatic patients and only two haplotypes were observed in the samples from asymptomatic cases. The findings of the current study showed that the  Leishmania  ITS1 from symptomatic VL and asymptomatic cases has significant genetic differences. Besides, infection with  Crithidia fasciculata  was reported, for the first time, in an asymptomatic case, which deserves further study.

Leishmania ITS1 Is Genetically Divergent in Asymptomatic and Symptomatic Visceral Leishmaniasis: Results of a Study in Southern Iran