Sequence analysis of mitochondrial co1 and nd1 genes in Echinococcus granulosus in Yunnan Province

doi:10.12140/j.issn.1000-7423.2023.03.007

Abstract

Abstract:

Objective To analyze the genotype and sequence polymorphisms of mitochondrial cytochrome oxidase 1（co1）and NADPH dehydrogenase subunit 1 (nd1) in Echinococcus granulosus from Yunnan Province. Methods Echinococcus of animal origin was collected from cysts isolated from liver or lung lesions of cattle, sheep and free-range pigs at slaughterhouses in Shangri-La, Daguan, Eryuan, Lushui and Weixi counties of Yunnan Province. Echinococcus of human origin was obtained from the focal tissues surgically removed from hospitals in Jianchuan, Yunlong, Longyang and Yulong Counties, then the E. granulosus was selected after pathogenic identification, and used to extract genomic DNA with a DNA extraction kit, followed by PCR amplification and sequencing of co1 and nd1 genes. The sequence homology and single nucleotide polymorphisms were analyzed using BLAST aligment. Phylogenetic trees based on co1 and nd1 were constructed by the neighbor-joining method using MEGA-X software. Results A total of 62 samples of Echinococcus were collected, 36 of which were pathogenically confirmed as E. granulosus. A total of 32 gene co1 sequences were successfully sequenced, of which 15 were type G1 and 17 were type G5, with a length of 824 bp and 807 bp, respectively. After submitting to GenBank database, the login numbers obtained were OP413393-OP413402, OP413498-OP413506 and OP420520. A total of 34 nd1 genes were sequenced successfully, of which 11 were type G1 and 23 were type G5, with lengths of 882 bp and 888 bp, respectively. The entry numbers obtained by submitting the GenBank database were OP471626-OP471638. The results of sequence polymorphism analysis showed that 1.94% (16/824) mutation sites in the co1 type G1 sequence and 3.10% (25/807) mutation sites in the G5 type sequence were found. In the nd1, 0.45% (4/882) and 2.93% (26/888) mutation sites were found in the type G1 sequence and the G5 type sequence, respectively. The results of multiple genotypic sequence comparison showed that there were 6 homologous sequences of the co1 gene, 4 of which were type G1, the genetic distances of homologous sequences were 0.000 0-0.001 1, 0.000 0-0.000 6, and 0.000 0-0.001 7, respectively. Also included two G5 types. The genetic distances of homologous sequences were 0.000 0-0.002 5 and 0.000 0-0.001 2, respectively. There were 3 homologous sequences in nd1 gene, of which 1 was type G1, and the genetic distance was 0.000 0-0.001 1, 2 of which were type G5, genetic distance was 0.000 0 and 0.000 0-0.001 1. Phylogenetic tree analysis showed that the phylogenetic tree based on co1 and nd1 genes formed two branches of type G1 and type G5, type G1 was closely related to the gene sequences of central and western China (Qinghai, Ningxia, Gansu, Sichuan), Bhutan, Iran and Jordan in the Middle East (GenBank entry number AF297617.1, KJ628328.1, EU072106.1, MW138946.1, AB688602.1). The G5 Type was closely related to Vietnam, China's Guangxi, the United Kingdom, Poland, Zambia, France, Pakistan, Brazil, Kenya and Namibia (GenBank login numbers MW558412.1, MN058591.1, KU378107.1, MZ322608.1, KU743915.1, KU743919.1, MN886291.1, KX010903.1, KU743918.1 and KX138068.1). Conclusion The prevalent isolate of E. granulosus in Yunnan Province are of genotype G1 and G5, and both co1 and nd1 gene reveals single nucleotide polymorphisms.

Key words: Echinococcus granulosus, co1 gene, nd1 gene, Sequence analysis, Yunnan Province

CLC Number:

R383.33

LI Benfu, WANG Zhengqing, XU Qian, ZI Jinrong, YAN Xinliu, PENG Jia, LI Jianxiong, CAI Xuan, WU Fangwei, YANG Yaming. Sequence analysis of mitochondrial co1 and nd1 genes in Echinococcus granulosus in Yunnan Province[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2023, 41(3): 306-311.

Figures/Tables 2

References 21

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