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Original Article

Genetic Diversity and Phylogenetic Analysis of the Iranian Leishmania Parasites Based on HSP70 Gene PCR-RFLP and Sequence Analysis

Sara Nemati1, Asghar Fazaeli1,*, Homa Hajjaran2, Ali Khamesipour3, Mohsen Falahati Anbaran4, Arezoo Bozorgomid2, Fatah Zarei5
The Korean Journal of Parasitology 2017;55(4):367-374.
Published online: August 31, 2017

1Department of Medical Parasitology and Mycology, School of Medicine, Zanjan University of Medical Sciences, Zanjan 4515613191, Iran

2Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran 14155-6446, Iran

3Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran 1416613675, Iran

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*Corresponding author (fazaeli@zums.ac.ir; fazaeli2017@gmail.com)
• Received: March 19, 2017   • Revised: May 5, 2017   • Accepted: May 23, 2017

Copyright © 2017 by The Korean Society for Parasitology and Tropical Medicine

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Genetic Diversity and Phylogenetic Analysis of the Iranian Leishmania Parasites Based on HSP70 Gene PCR-RFLP and Sequence Analysis
Korean J Parasitol. 2017;55(4):367-374.   Published online August 31, 2017
DOI: https://doi.org/10.3347/kjp.2017.55.4.367
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Genetic Diversity and Phylogenetic Analysis of the Iranian Leishmania Parasites Based on HSP70 Gene PCR-RFLP and Sequence Analysis
Korean J Parasitol. 2017;55(4):367-374.   Published online August 31, 2017
DOI: https://doi.org/10.3347/kjp.2017.55.4.367
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Genetic Diversity and Phylogenetic Analysis of the Iranian Leishmania Parasites Based on HSP70 Gene PCR-RFLP and Sequence Analysis
Image Image Image
Fig. 1 Map of Iran, showing our sampling localities. Locality numbers are the same as in Table 1.
Fig. 2 Agarose gel (3%) showing 3 HSP70 PCR-RFLP profiles for Leishmania reference strains after digestion with HaeIII. Lane1, L. tropica; lane 2, L. tropica; lane 3, L. major; lane 4, L. major, lane 5, L. infantum; lane 6, L. infantum; lane M, 100-bp size marker.
Fig. 3 ML tree summarizing the relationship between HSP70 sequences of Leishmania species. Bootstrap resampling values are provided at each fork (given only if ≥40%). Iranian isolates are represented by circle (L. major), triangle (L. tropica), and square (L. infantum) symbols.

Fig. 1

Fig. 2

Fig. 3

Genetic Diversity and Phylogenetic Analysis of the Iranian Leishmania Parasites Based on HSP70 Gene PCR-RFLP and Sequence Analysis

Details of the original and GenBank Leishmania isolates used in the present study

No.a Sample ID Species Localityb Host Disease Sample ID Species Localityc
1 LM4 L. major Esfahan Human CL JN628986.1 L. infantum Georgia
2 LM5 L. major Ardestan Human CL JN676923 L. infantum Georgia
3 LM6 L. major Mashhad Human CL JX021443 L. turanica China
4 LM7 L. major Sistan Baluchestan Human CL JX021433 L. infantum China
5 LM8 L. major Shiraz Human CL JX021432 L. infantum China
6 LM10 L. major Agha Aliabas Human CL FN395025 L. tropica India
1 LM12 L. major Esfahana Human CL FN395026 L. tropica Kenya
5 LM13 L. major Shiraz Human CL FN395022 L. major Sudan
5 LM14 L. major Shiraz Human CL FN395023 L. major Israil
7 LM28 L. major Dehloran Human CL FN395024 L. major Kenya
8 LM29 L. major Ilam Human CL XM001684512 L. major Israel
8 LM30 L. major Ilam Human CL FN395031 L. infantum Malta
9 LM31 L. major Golestan Human CL FN395032 L. infantum Portugal
10 LM36 L. major Bandar_Abbas Human CL FN395033 L. infantum Malta
11 LT27 L. major Kermanshah Human CL XM001470287 L. infantum Spain
3 LT L. tropica Mashhad Human CL GU071173 L. braziliensis Brazil
12 LT2 L. tropica Bam Human CL KG905366 L. donovani India
12 LT3 L. tropica Bam Human CL GU071178 L. guyanensis Brazil
3 LT16 L. tropica Mashhad Human CL HF586354 L. amazonensis Panama
17 LT9 L. tropica Afghanistan Human CL HF586353 L. amazonensis Colombia
17 LT19 L. tropica Afghanistan Human CL JX312712 L. donovani China
17 LT21 L. tropica Afghanistan Human CL FN669773 L. donovani Ethiopia
9 LT22 L. tropica Golestan Human CL GU071174 L. lainsoni Brazil
3 LT25 L. tropica Mashhad Human CL HF586413 L. mexicana Ecuador
5 LT26 L. tropica Shiraz Human CL LN907842 L. mexicana Ecuador
13 L.VT5 L. tropica Ardebil Human CL FN395056 L. naiffi Brazil
14 L.VT6 L. tropica Tehran Human CL FN395055 L. panamensis Panama
15 LI.1 L. infantum Meshkin Shahr Canine VL HF586367 L. panamensis Panama
15 LI.2 L. infantum Meshkin Shahr Canine VL HF586368 L. peruviana Peru
15 LI.3 L. infantum Meshkin Shahr Canine VL FN395044 L. peruviana Peru
15 LI.7 L. infantum Meshkin Shahr Canine VL GU071175 L. shawi Brazil
15 LI.8 L. infantum Meshkin Shahr Canine VL HF586356 L. turanica China
16 L4 L. major Hamedan Rodent JX021442 L. turanica China
16 L5 L. major Hamedan Rodent FN395037 L. chagasi Brazil
16 L38 L. major Hamedan Rodent FN395036 L. chagasi Brazil
16 L40 L.major Hamedan Rodent FN395035 L. chagasi Brazil
  JN628988 L. infantum Georgiac   HF586355 L. gerbilli China
  JN628987 L. infantum Georgiac   EF108422 T. rangeli Honduras

aLocality number on Fig. 1.

bOriginal Leishmania strains from different districts of Iran.

cStrains studied by other researchers, taken from NCBI.

Basic parameters of genetic diversity and neutrality tests, for 3 Iranian Leishmania species

Species Diversity parameters Neutrality tests
Na Sb Hc Parsimony informative sites Singleton variable sites h±SDd πe Tajima’s D Fu’s Fs
L. major 20 10 3 1 9 0.195±0.11 0.00093 −2.14f 1.42
L. tropica 11 2 2 2 0 0.436±0.13 0.00075 0.85 2.01
L. infantum 5 0 1 0 0 0.00±0.00 0.0000 N.A.g N.A.

aNo. of sequences.

bNo. of polymorphic sites.

cNo. of haplotypes.

dHaplotype diversity (h±SD).

eNucleotide diversity.

fP<0.05.

gNot available.

Table 1 Details of the original and GenBank Leishmania isolates used in the present study

Locality number on Fig. 1.

Original Leishmania strains from different districts of Iran.

Strains studied by other researchers, taken from NCBI.

Table 2 Basic parameters of genetic diversity and neutrality tests, for 3 Iranian Leishmania species

No. of sequences.

No. of polymorphic sites.

No. of haplotypes.

Haplotype diversity (h±SD).

Nucleotide diversity.

P<0.05.

Not available.

Table 1

Table 2

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