论文部分内容阅读
目的 对卵形疟原虫（Plasmodium ovale）curtisi亚种和wallikeri亚种的编码细胞色素b（Cytb），细胞色素c氧化酶 I(Cox I)和乳酸脱氢酶（LDH）基因的单核苷酸多态性进行分析。 方法 选择浙江省疟疾诊断参比实验室2011-2014年确诊的卵形疟原虫感染患者血样，提取卵形疟原虫DNA，巢式PCR扩增卵形疟原虫Cytb、Cox I和LDH基因的部分片段，对扩增产物进行双向测序。测序序列经校对、拼接后，应用MEGA软件对测序序列和相应的氨基酸序列进行比对分析，发现单核苷酸多态性位点以及氨基酸突变位点。 结果 19例卵形疟原虫感染者（其中curtisi亚种11例，wallikeri亚种8例）均为赴非洲返国人员。PCR扩增Cytb、Cox I和LDH基因的产物长度分别约为735、1 323和355 bp。curtisi和wallikeri两个亚种在Cytb、Cox I和LDH基因分别发现10、13、13个单核苷酸二态性位点，且大多数的二态性位点属于同义突变。此外，两个亚种内部也存在核苷酸多态性，同时发现1例血样的Cox I基因存在693个碱基的缺失。 结论 对卵形疟患者血样的Cytb、Cox I和LDH基因进行了单核苷酸多态性分析，丰富了卵形疟原虫基因多态性信息。
OBJECTIVE: To characterize the cytochrome b (Cytb), cytochrome c oxidase I (Cox I) and lactate dehydrogenase (LDH) genes of Plasmodium ovale subsp. Curtisi and wallikeri subsp. Polymorphism analysis. Methods Blood samples were collected from malaria-infected malaria patients in Zhejiang Reference Laboratory for Diagnosis of Malaria from 2011 to 2014. Plasmodium falciparum DNA was extracted. Nested PCR was used to amplify some fragments of Cytb, Cox I and LDH genes of Plasmodium ovalensis , The amplified products were bi-directionally sequenced. Sequencing and alignment of the sequencing sequence, MEGA software was used to analyze the sequence and the corresponding amino acid sequence, and single nucleotide polymorphism and amino acid mutation sites were found. Results 19 cases of Plasmodium ovale infection (including 11 curtisi subspecies and 8 wallikeri subspecies) were returned to Africa. The product lengths of Cytb, Cox I and LDH genes amplified by PCR were about 735, 1323 and 355 bp, respectively. Two subspecies curtisi and wallikeri found 10, 13 and 13 single nucleotide polymorphisms in Cytb, Cox I and LDH genes, respectively, and most of the two polymorphic sites belong to synonymous mutations. In addition, nucleotide polymorphisms were also found in both subspecies. One sample of blood was found to have a 693-bp deletion of the Cox I gene. Conclusion Single nucleotide polymorphisms (SNPs) of Cytb, Cox I and LDH genes in blood samples from oval-shaped malaria patients were analyzed, which enriched the genetic information of Plasmodium ovale.