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目的:测定恶性疟原虫中国海南分离株MSP1 基因的全序列,并进行多态性分析。方法:来自海南省保亭县2 例恶性疟患者的血样(分别滴在滤纸上)直接制备基因组DNA 后,用MSP1 基因特异的5 对寡核苷酸引物在体外扩增目的基因,用ABIPRISMTM末端标记循环测序试剂盒进行直接测序,并将测序结果与已知的等位基因型进行比较分析。结果:获得了2 个恶性疟原虫中国分离株MSP1 基因的全序列,与已知的MSP1 等位基因型进行序列比较,证实其属于MAD20 型。推导其氨基酸序列除了第2 区、第4 区和第8 区以外,其余序列完全一致。其中HN2 的第4 区含K1 型序列。结论:两株恶性疟原虫海南株的MSP1 基因属于MAD20 型,与MAD20 等位基因比较,其推导的氨基酸序列存在一些小差异。本文首次提供了恶性疟原虫中国(海南)分离株MSP1 基因多态性的依据
Objective: To determine the complete sequence of MSP1 gene of Plasmodium falciparum isolates from Hainan in China and analyze its polymorphism. Methods: Two genomic DNAs were prepared directly from two blood samples of falciparum malaria patients from Baoting, Hainan Province. The target genes were amplified in vitro by using five pairs of oligonucleotide primers specific to MSP1 gene. The end of ABIPRISMTM Marker cycle sequencing kit for direct sequencing, and the sequencing results were compared with the known alleles. Results: The complete sequence of MSP1 gene of two Chinese Plasmodium falciparum isolates was obtained. Compared with the known MSP1 allele, it was confirmed that it belonged to MAD20 type. The deduced amino acid sequence except for region 2, region 4 and region 8, the other sequences are exactly the same. The HN2 region 4 contains K1 type sequence. CONCLUSION: The MSP1 gene of two strains of Plasmodium falciparum in Hainan belongs to MAD20 type. Compared with the MAD20 allele, the deduced amino acid sequence of MSP1 gene is slightly different. This article for the first time provides a basis for the genetic polymorphism of the MSP1 gene of P. falciparum isolates from China (Hainan)