Abstract
Objective To compare the difference between the analysis on the phylogenetic relationships of Leishmania isolates from different foci in China with partial fragment of small subunit ribosomal RNA (SSUrRNA) gene sequences or kinetoplast cytochrome oxidase Ⅱ (COX Ⅱ) gene sequences.Methods Genomic DNA and kinetoplast DNA were extracted from the promastigotes.The CO Ⅱ and SSUrRNA genes were amplified using respective primers.Products of PCR were sent to a commercial company for sequencing using the same PCR primer.Combined sequences matrix was aligned using Clustal X (1.8) with default settings and refined manually.Bayesian tree was constructed using Mrbayes-3.1.2.Results Leishmania isolates from different foci in China does not form a mono-phyletic group.The isolate MHOM/CN/93/GS7 and the isolate IPHL/CN/77/XJ771 belong to Leishmania donovani complex.Six haplotypes representing 10 Chinese Leishmania isolates(except isolates MHOM/CN/93/GS7,IPHL/CN/77/XJ771,MHOM/CN/84/JS1 and MGER/CN/60/GSGER20) form a monophyletic group (results from CO Ⅱ genes database).Eleven Chinese Leishmania isolates (except IPHL/CN/77/XJ771,MHOM/CN/93/GS7,MRHO/CN/88/KX G-2,MHOM/CN/84/JS1,MGER/CN/60/GSGER20) form a subclade,near to Leishmania tarentolae (results from SSUrRNA genes database); the isolate MHOM/CN/84/JS1 is sister of L.tropica representing L.tropica complex with high posterior probability (PP=1.00),instead of clustering with L,donovani complex or with other 10 Chinese isolates.Conclusions The analysis results with either SSUrRNA or COX Ⅱ seemed consistent.However kinetoplast cytochrome oxidase Ⅱ gene might be a more reliable genetic marker than SSUrRNA to analyze the phylogentic relationship of Chinese Leishmania isolates. Key words: Chinese Leishmania isolates; Promastigote; Phylogeny; Small subunit ribosomal RNA; Kinetoplast cytochrome oxidase Ⅱ
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