Molecular characterization of Leishamania isolates from China by inter-simple sequence repeat polymerase chain reaction

Abstract

Leishmania has distinct epidemiological and biological characteristics and causes a variety of clinical symptoms. To understand the genetic diversity and the phylogenetic relationships among Leishmania isolates from China, 29 Leishmania isolates from different geographic origins, vectors, and hosts were analyzed using 21 inter-simple sequence repeat polymerase chain reaction (ISSR-PCR) primers. A total of 864 polymorphic bands were obtained. According to the results of the neighbor-joining phylogenetic tree and principal component analysis, the 29 isolates studied clustered into six groups. Isolates of Leishmania donovani complex from China share the highest similarity with the reference strain of L. donovani (DD8). This study helps to elucidate the genetic relationship among Leishmania isolates from China and similarities between Chinese isolates and World Health Organization reference strains. Furthermore, ISSR-PCR could also be a quick, simple, and reliable method for Leishmania species identification.

Phylogenetic and Evolutionary Analysis of Chinese Leishmania Isolates Based on Multilocus Sequence Typing

“…The result of seven conserved genes indicated that the five strains were closely related to strain 3720, which was classified as L. turanica . Our results support the conclusion of previous studies [29] that these five strains were grouped as L. turanica . Two strains (SELF-7 and EJNI-154) in clade C have been previously identified as L. gerbilli [29].…”

“…Clade A was further divided into two subpopulations, clade A1 and clade A2. Five Chinese strains formed a group (clade B) with WHO L. turanica reference strain 3720, as reported previously [29]. Two distinct groups were also produced, one including SELF-7 and EJNI-154 (clade C) which were closely related to clade B in the tree, and the other population (clade D), consisting of LIZRD and KXG-E, was separated from the other Chinese isolates.…”

“…The first group included 13 Chinese isolates and a WHO L. donovani reference strain DD8. The second group included five Chinese isolates (strain KXG-Y, KXG-R, KXG-57, KXG-11 and QITAI-15) and a WHO L. turanica reference strain 3720 as reported in a previous study [29]. When the two groups were compared, several nucleotide differences at numerous sites were found, so the DNA sequences were analyzed for the two groups, respectively.…”

“…Two strains, MCAN/CN/11/1101 and MCAN/CN/11/1102, were isolated from two dogs in China in 2011. The other 26 strains included in this study were kindly provided by Professor Wang Junyun at the Shanghai Municipal Center for Disease Control and Prevention, and has been previously analyzed by Wang Y et al [29]. For the isolation of MCAN/CN/11/1101 and MCAN/CN/11/1102, oral informed consent was obtained from the guardians of the two dogs.…”

Phylogenetic analysis of HSP70 and cyt b gene sequences for Chinese Leishmania isolates and ultrastructural characteristics of Chinese Leishmania sp.

“…Meanwhile, we found that the strains KXG-XU and KXG-LIU (HSP70 gene sequence JX021433 and JX021432), isolated from CL patients in Karamay, Xinjiang, also belong to L. donovani according to HSP70 BI tree, which was in accord with the study based on ISSR-PCR by Wang et al (2010b). Accordingly, since the five strains KXG-918, KXG-927, KXG-65, KXG-XU, and KXG-LIU were all isolated in Karamay, Xinjiang, and grouped together based on HSP70 data, it can be inferred that L. donovani is the pathogen of CL in Karamay and Phlebotomus major wui is the vector.…”