Objective Human Lyme Borreliosis （LB）, which is caused by Borrefia burgdorferi sensu lato （B. burgdorferi）, has been identified as a major arthropod-borne infectious disease in China. We aimed to develop a multipl...Objective Human Lyme Borreliosis （LB）, which is caused by Borrefia burgdorferi sensu lato （B. burgdorferi）, has been identified as a major arthropod-borne infectious disease in China. We aimed to develop a multiple locus variable-number tandem repeat （VNTR） analysis （MLVA） assay for the genotyping of Borrelia burgdorJ：eri strains detected in China. Methods B. garinii PBi complete 904.246 kb chromosome and two plasmids （cp26 and Ip54） were screened by using Tandem Repeats Finder program for getting potential VNTR loci, the potential VNTR loci were analyzed and identified with PCR and the VNTR loci data were analyzed and MLVA clustering tree were constrcted by using the categorical coefficient and the unweighted pair-group method with arithmetic means （UPGMA）. Results We identified 5 new VNTR loci through analyzing 47 potential VNTR loci. We used the MLVA protocol to analyse 101 B. burgdorferi strains detected in China and finally identified 51 unique genotypes in 4 major clusters including B. burgdorferi sensu stricto （B.b.s.s）, B. garinii, B. a[zelii, and B. valaisiana, consistent with the current MLSA phylogeny studies. The allele numbers of VNTR-1, VNTR-2, VNTR-3, VNTR-4, and VNTR-5 were 7, 3, 9, 7, and 6. The Hunter-Gaston index （HGI） of five VNTR loci were 0.79, 0.22, 0.77, 0.71, and 0.67, respectively. The combined HGI of five VNTR loci was 0.96. Clustering of the strains of Xinjiang, Inner Mongolia and Heilongjiang was confirmed, and this situation was consistent with the close geographical distribution of those provinces. Conclusion The MLVA protocol esytablished in this study is easy and can show strains＇ phylogenetic relationships to distinguish the strains of Borrelia species. It is useful for further phylogenetic and epidemiological analyses of Borrelia strains.展开更多
Objective To study the polymorphism in P66 and its human B-cell epitopes of Borrelia burgdorferi strains in China.Methods Polymerase chain reaction(PCR)and sequencing were used to obtain the P66 sequences of59 Chinese...Objective To study the polymorphism in P66 and its human B-cell epitopes of Borrelia burgdorferi strains in China.Methods Polymerase chain reaction(PCR)and sequencing were used to obtain the P66 sequences of59 Chinese B.burgdorferi.Then the sequences were analyzed by MEGA 5.10 software and compared with the human B-cell epitope sequences from the Immune Epitope Database(IEDB)based on the reference strain of each genotype.Results Results showed that genetic and amino acid diversity presented in the 66 kD protein of all 59 Chinese strains,especially in Borrelia garinii(B.g)and Borrelia afzelii(B.a)strains.B.g strains were divided into three subclusters and two scattered strains JC1-7 and JC2-2 according to the amino acid sequences of P66.The P66 sequences of 15 Xinjiang strains represented by XI91-12 in the B.g subcluster1,changed from CAA to TAA codon at 508 aa position,resulting in early termination.Bases A and C were inserted at sequence position 1523 bp of strains FP1,LB20,LB21,and SZ21 in the B.a genotype,which resulted to early termination at position 511 aa.G base was inserted at 438 bp of LIP94-11 strain,which led to early termination at position 172 aa.Conclusion In P66 of 59 Chinese strains,polymorphisms were widely distributed.More importantly,the P66 amino acid sequences of B.g strains had a certain regional character.One of the characteristics of Xinjiang B.g isolates might be the variation at the 508 aa location in 15 Xinjiang B.g strains,which may be related to the strains’pathogenicity in this area.展开更多
目的研究我国伯氏疏螺旋体(Borrelia burgdorferi sensu lato),即莱姆病螺旋体菌株中bmpA基因及其人B细胞表位区的序列多态性。方法选取61株莱姆病螺旋体分离株,PCR扩增其bmpA基因并测序后,结合4种基因型参考菌株序列,与从免疫表位数据...目的研究我国伯氏疏螺旋体(Borrelia burgdorferi sensu lato),即莱姆病螺旋体菌株中bmpA基因及其人B细胞表位区的序列多态性。方法选取61株莱姆病螺旋体分离株,PCR扩增其bmpA基因并测序后,结合4种基因型参考菌株序列,与从免疫表位数据库(Immune Epitope Database,IEDB)中查询到的B细胞表位序列进行比对分析。结果中国B.garinii基因型菌株的cluster2、cluster3以及B.afzelii和B.valaisiana基因型所有菌株与参考序列比对后发现分别有3、9、5和18个异义突变。B细胞表位改变主要发生在B.afzelii基因型全部分离株和B.garinii基因型的cluster3及5株非成簇菌株。B.afzelii基因型菌株在B细胞表位区的异义突变A125S影响了2个B细胞表位;B.garinii基因型的cluster3和5株非成簇菌株B细胞表位区的3个异义突变导致5个B细胞表位均发生改变。结论中国莱姆病螺旋体bmpA基因在4种基因型间和B.garinii基因型内存在较大多态性。展开更多
基金supported by Natural Science foundation(Grant No.31100105)China Mega-Project for Infectious Disease(2012ZX10004-215 and 2013ZX10004221)
文摘Objective Human Lyme Borreliosis （LB）, which is caused by Borrefia burgdorferi sensu lato （B. burgdorferi）, has been identified as a major arthropod-borne infectious disease in China. We aimed to develop a multiple locus variable-number tandem repeat （VNTR） analysis （MLVA） assay for the genotyping of Borrelia burgdorJ：eri strains detected in China. Methods B. garinii PBi complete 904.246 kb chromosome and two plasmids （cp26 and Ip54） were screened by using Tandem Repeats Finder program for getting potential VNTR loci, the potential VNTR loci were analyzed and identified with PCR and the VNTR loci data were analyzed and MLVA clustering tree were constrcted by using the categorical coefficient and the unweighted pair-group method with arithmetic means （UPGMA）. Results We identified 5 new VNTR loci through analyzing 47 potential VNTR loci. We used the MLVA protocol to analyse 101 B. burgdorferi strains detected in China and finally identified 51 unique genotypes in 4 major clusters including B. burgdorferi sensu stricto （B.b.s.s）, B. garinii, B. a[zelii, and B. valaisiana, consistent with the current MLSA phylogeny studies. The allele numbers of VNTR-1, VNTR-2, VNTR-3, VNTR-4, and VNTR-5 were 7, 3, 9, 7, and 6. The Hunter-Gaston index （HGI） of five VNTR loci were 0.79, 0.22, 0.77, 0.71, and 0.67, respectively. The combined HGI of five VNTR loci was 0.96. Clustering of the strains of Xinjiang, Inner Mongolia and Heilongjiang was confirmed, and this situation was consistent with the close geographical distribution of those provinces. Conclusion The MLVA protocol esytablished in this study is easy and can show strains＇ phylogenetic relationships to distinguish the strains of Borrelia species. It is useful for further phylogenetic and epidemiological analyses of Borrelia strains.
基金supported by Major Projects of the thirteenth Five Year Special for infectious diseases[2016ZX10004001-004 and 2018ZX10714002]。
文摘Objective To study the polymorphism in P66 and its human B-cell epitopes of Borrelia burgdorferi strains in China.Methods Polymerase chain reaction(PCR)and sequencing were used to obtain the P66 sequences of59 Chinese B.burgdorferi.Then the sequences were analyzed by MEGA 5.10 software and compared with the human B-cell epitope sequences from the Immune Epitope Database(IEDB)based on the reference strain of each genotype.Results Results showed that genetic and amino acid diversity presented in the 66 kD protein of all 59 Chinese strains,especially in Borrelia garinii(B.g)and Borrelia afzelii(B.a)strains.B.g strains were divided into three subclusters and two scattered strains JC1-7 and JC2-2 according to the amino acid sequences of P66.The P66 sequences of 15 Xinjiang strains represented by XI91-12 in the B.g subcluster1,changed from CAA to TAA codon at 508 aa position,resulting in early termination.Bases A and C were inserted at sequence position 1523 bp of strains FP1,LB20,LB21,and SZ21 in the B.a genotype,which resulted to early termination at position 511 aa.G base was inserted at 438 bp of LIP94-11 strain,which led to early termination at position 172 aa.Conclusion In P66 of 59 Chinese strains,polymorphisms were widely distributed.More importantly,the P66 amino acid sequences of B.g strains had a certain regional character.One of the characteristics of Xinjiang B.g isolates might be the variation at the 508 aa location in 15 Xinjiang B.g strains,which may be related to the strains’pathogenicity in this area.
