中缅边境及泰国东南地区恶性疟原虫K13基因侧翼微卫星多态性研究
本文选题:恶性疟原虫 + 微卫星 ; 参考:《中国寄生虫学与寄生虫病杂志》2017年03期
【摘要】:目的研究中缅边境拉咱地区和泰国东南Srisaket、Trat和Ranong等3个地区恶性疟原虫(Plasmodium falciparum)K13基因侧翼微卫星的多态性。方法取前期经基因型研究的恶性疟原虫样品42份,其中中缅边境拉咱地区22份(K13基因R539T突变型和野生型各11份),泰国3个地区20份(K13基因R539T突变型9份和野生型11份),对上述样品K13基因上游31 900、6 360、150 bp和下游1 700、11 000、31 500 bp共6个位点的微卫星片段进行PCR扩增,PCR产物经毛细管电泳仪检测片段长度,并采用Bio Calculator软件进行数据分析,Gen ALEx软件计算等位基因频率和期望杂合度(He)。结果中缅边境拉咱地区和泰国3个地区恶性疟原虫虫株K13基因两侧的6个微卫星位点共检测到46个等位基因,在上游6 360 bp位点的等位基因长度分别集中在286和278 bp,等位基因频率为73%和67%。上游31 900 bp位点等位基因长度分别集中在211和205 bp,等位基因频率为55%和44%。上游150 bp及下游1 700 bp位点等位基因长度主要集中在195和209 bp,其他两个位点未出现集中分布的情况。中缅边境拉咱地区虫株的单倍体型为H1至H4型,泰国3个地区虫株除了2株H2单体型外,其余均为H5和H6型。42份样品的6个微卫星位点的平均He分别为0.49±0.23、0.42±0.33和0.72±0.12、0.67±0.11。R539T突变的虫株在毗邻K13基因的微卫星位点处He明显降低,在上游150 bp处分别为0.17和0,在下游1 700 bp处分别为0.3和0。结论中缅边境拉咱地区及泰国3个地区K13基因为R539T突变型的恶性疟原虫虫株均受到青蒿素选择压力,等位基因型在上游6 360和31 900 bp两个位点上具有明显的差异。
[Abstract]:Objective to study the polymorphism of Plasmodium falciparum)K13 gene flanking microsatellite in Laasan region of Sino-Myanmar border and Srisakettrat and Ranong in southeast Thailand. Methods 42 samples of Plasmodium falciparum were collected from genotypic study. There were 22 R539T mutant and 11 wild type in R539T gene, 9 in R539T mutation and 11 in wild type in three regions of Thailand. The upstream of K13 gene was 31,900-6 360150 BP and the downstream of K13 gene was 1,7000.11. The fragment length was detected by capillary electrophoresis after PCR amplification of the microsatellite fragment with a total of 6 loci (31,500 BP), and the length of the fragment was determined by capillary electrophoresis. The allele frequency and expected heterozygosity were calculated by Bio Calculator software. Results A total of 46 alleles were detected at 6 microsatellite loci on both sides of the K13 gene of Plasmodium falciparum strain in Laasan region of the Sino-Myanmar border and 3 regions in Thailand. The allele length of the 6360 BP locus in the upstream was 286 and 278 BP, respectively. The allele frequencies were 73% and 67%, respectively. The alleles at 31,900 BP loci were located at 211 BP and 205 BP, respectively. The allele frequencies were 55% and 44 BP, respectively. The alleles of 150 BP upstream and 1 700 BP downstream were mainly located at 195 and 209 BP, while the other two loci were not concentrated. H1-H4 haploid strains were found in Laasan area of Sino-Myanmar border, and two H _ 2 haplotypes were found in three strains in Thailand. The average he of the 6 microsatellite loci in the other samples were 0.49 卤0.230.42 卤0.33 and 0.72 卤0.120.67 卤0.11.R539T, respectively, and the he at the microsatellite site adjacent to the K13 gene was significantly decreased. It was 0.17 and 0 at 150 BP upstream and 0.3 and 0 at 1 700 BP downstream, respectively. Conclusion the K13 gene of R539T mutant strain of Plasmodium falciparum is under artemisinin selection pressure in Laasan region and three regions of Thailand, and the alleles are different at 6360 BP and 31,900 BP loci upstream.
【作者单位】: 第二军医大学热带传染病学教研室;
【基金】:国家自然科学基金(No.81220108019)~~
【分类号】:R382.31
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