CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >

2020 , Vol. 38 >Issue 2: 181 - 187

DOI: https://doi.org/10.12140/j.issn.1000-7423.2020.02.008

ORIGNAL ARTICLES

Evaluation of the value of K26 sequence applied in identification of Leishmania isolates in China

  • Jian-xiu LIU ,
  • Chun-hua GAO ,
  • Yue-tao YANG ,
  • Bin ZHENG ,
  • Jun-yun WANG
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  • National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; WHO Collaborating Centre for Tropical Diseases; National Center for International Research on Tropical Diseases, Ministry of Science and Technology; Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai 200025, China

Received date: 2019-12-02

  Online published: 2020-05-11

Supported by

Supported by the National Science and Technology Major Program of China(2018ZX10734-404);the National Natural Science Foundation of China(81472923)

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Abstract

Objective To evaluate the application of Leishmania K26 sequence in identification of Leishmania isolates in China.Methods Sixteen Leishmania isolates collected from different domestic endemic areas and 6 international reference strains were cultured for collection of promastigotes, from which DNA was extracted for amplification of K26 sequence by PCR. The K26 sequence was confirmed by sequencing. The sequence alignment was made using the ClustalX2 software. A phylogenetic tree was constructed based on the K26 sequences of highly-homologous Leishmania isolates downloaded from the GenBank database, to analyze the genetic relationship among the Leishmania isolates from different endemic areas in China.Results PCR amplification showed a specific band from all the 16 isolates and the DD8 international reference strain, while no specific band was found from five non-L. donovani complex strains. Sequence analysis indicates that all 3 isolates 801, KS-2 and KS-6 from the endemic areas of anthroponotic leishmaniasis generated a 920 bp fragment in PCR; 4 isolates XJ771, JIASHI-1, JIASHI-2 and JIASHI-5 from endemic areas of natural-sourced leishmaniasis all generated a 491 bp fragment in amplification reaction; 4 strains Cy, SC6, Hebei-Lv and Shanxi-Yang from endemic areas of zoonosis (specifically canine) leishmaniasis generated a 404 bp in amplification; 5 strains KXG-Xu, KXG-Liu, KXG-65, KXG-918 and KXG-927 from endemic areas of cutaneous leishmaniasis all generated a 449 bp fragment in amplification. Among the endemic areas, the K26 sequences of intra-area isolates showed 100% homology, while the sequences of enter-area isolates displayed a lower homology (40.2%-91.4%). Phylogenetic tree analysis showed that the 16 isolates of Leishmania in China clustered into 2 groups and 3 subgroups, among which nine isolates including KXG-Xu, KXG-Liu, KXG-65, KXG-918, KXG-927, XJ771, JIASHI-1, JIASHI-2, and JIASHI-5 clustered into subgroup A, while five isolates Cy, SC6, Hebei-Lv, Shanxi-Yang and L. infantum clustered into subgroup B, and both subgroups clustered into groupⅠ. The 801, KS-2 and KS-6 isolates clustered with L. donovani as subgroup C, which further clustered with the DD8 international reference isolate into group Ⅱ.Conclusion The K26 sequences of Leishmania can be used for identification of Leishmania isolates in China.

Key words： Leishmaniasis; Leishmania; Isolate; K26; Phylogenetic tree; China

Cite this article

Jian-xiu LIU , Chun-hua GAO , Yue-tao YANG , Bin ZHENG , Jun-yun WANG . Evaluation of the value of K26 sequence applied in identification of Leishmania isolates in China[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2020 , 38(2) : 181 -187 . DOI: 10.12140/j.issn.1000-7423.2020.02.008

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