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目的分析山东省间日疟原虫MSP-1和CSP基因类型及其同源性,为病例溯源提供科学依据。方法采集2011年山东省报告的12例间日疟患者血样,提取疟原虫基因组DNA;分别根据间日疟原虫MSP-1和CSP基因序列设计引物,进行巢式PCR扩增、酶切、测序、序列比对及同源性分析。结果 12份间日疟患者血样MSP-1基因全部出现470bp扩增条带以及350、120 bp酶切片段,均为Sal-1型;MSP-1进化树分析显示,9份省内感染者样品序列同属一个分枝,1份印度感染者样品序列与印度分离株位于同一分枝。12份间日疟患者样品CSP基因均包含GDRA(D/A)GQPA序列,为PV-Ⅰ型,其中10份省内感染者和1份广东感染者样品CSP基因出现560~840 bp和150~230 bp两种扩增条带,为PV-Ⅰ型温带族,1份在印度感染者样品CSP基因仅出现560~840 bp条带,为PV-Ⅰ型热带族。CSP进化树表明,10份省内感染者及1份广东感染者样品序列同属一个分枝,1份在印度感染者样品序列与印度和印度尼西亚分离株位于同一分枝。结论山东省本地感染间日疟原虫MSP-1基因型均为Sal-1型,CSP基因型均为PV-Ⅰ型温带族,本地虫株具有较强的基因同源性。
Objective To analyze the genotypes and homologies of MSP-1 and CSP genes of Plasmodium vivax in Shandong province and to provide a scientific basis for the follow-up of cases. Methods The blood samples of 12 malaria patients were collected from Shandong Province in 2011, and the genomic DNA of Plasmodium was extracted. Primers were designed according to MSP-1 and CSP gene sequence of Plasmodium vivax respectively, Sequence alignment and homology analysis. Results The MSP-1 gene in all the 12 samples of Plasmodium falciparum showed a 470 bp amplified fragment and a 350 bp fragment of 120 bp, all of which were type Sal-1. MSP-1 phylogenetic tree analysis showed that 9 samples The sequences belong to the same branch, and a sample of Indian Infected samples is in the same branch as the Indian isolate. The CSP gene of 12 samples of Plasmodium vivax contained GDRA (D / A) GQPA sequence, which was PV-Ⅰ type. The CSP gene in 10 infected persons and 1 infected person in Guangdong province showed 560 ~ 840 bp and 150 ~ 230 bp two kinds of amplification bands are PV-Ⅰ type temperate family, and 1 copy of the CSP gene in India showed only 560 ~ 840 bp band, which is type PV-Ⅰ tropical family. The CSP phylogenetic tree showed that the samples from 10 provinces and 1 infected Guangdong were all the same and 1 was located in the same branch with Indian and Indonesian isolates. Conclusion The MSP-1 genotypes of Plasmodium vivax in Shandong Province are both Sal-1 type and CSP genotypes are PV-Ⅰ type temperate family. The local strains have strong gene homology.