Kritsiriwuthinan Kanyanan, Ngrenngarmlert Warunee, Patrapuvich Rapatbhorn, Phuagthong Supaksajee, Choosang Kantima
Faculty of Medical Technology, Rangsit University, Pathumthani 12000, Thailand.
Department of Community Medical Technology, Faculty of Medical Technology, Mahidol University, Nakhon Pathom 73170, Thailand.
J Trop Med. 2023 Aug 11;2023:8855171. doi: 10.1155/2023/8855171. eCollection 2023.
Considering the importance of merozoite surface proteins (MSPs) as vaccine candidates, this study was conducted to investigate the polymorphism and genetic diversity of merozoite surface protein 3-alpha () in Thailand. To analyze genetic diversity, 118 blood samples containing were collected from four malaria-endemic areas in western and southern Thailand. The DNA was extracted and amplified for the gene using nested PCR. The PCR products were genotyped by PCR-RFLP with I and I restriction enzymes. The combination patterns of I and I RFLP were used to identify allelic variants. Genetic evaluation and phylogenic analysis were performed on 13 sequences, including 10 sequences from our study and 3 sequences from GenBank. The results revealed three major types of , 91.5% allelic type A (∼1.8 kb), 5.1% allelic type B (∼1.5 kb), and 3.4% allelic type C (∼1.2 kb), were detected based on PCR product size with different frequencies. Among all , 19 allelic subtypes with I RFLP patterns were distinguished and 6 allelic subtypes with I RFLP patterns were identified. Of these samples, 73 (61%) and 42 (35.6%) samples were defined as monoallelic subtype infection by I and I PCR-RFLP, respectively, whereas 77 (65.3%) samples were determined to be mixed-allelic subtype infection by the combination patterns of I and I RFLP. These results strongly indicate that gene is highly polymorphic, particularly in blood samples collected from the Thai-Myanmar border area (the western part of Thailand). The combination patterns I and I RFLP of the gene could be considered for use as molecular epidemiologic markers for genotyping isolates in Thailand.
鉴于裂殖子表面蛋白（MSPs）作为疫苗候选物的重要性，本研究旨在调查泰国裂殖子表面蛋白3-α（）的多态性和遗传多样性。为了分析遗传多样性，从泰国西部和南部四个疟疾流行地区收集了118份含有的血样。提取DNA并使用巢式PCR扩增基因。PCR产物用I和I限制性内切酶通过PCR-RFLP进行基因分型。I和I RFLP的组合模式用于鉴定等位基因变体。对13个序列进行了遗传评估和系统发育分析，其中包括我们研究中的10个序列和GenBank中的3个序列。结果显示，基于PCR产物大小检测到三种主要类型的，等位基因A类型（约1.8 kb）占91.5%，等位基因B类型（约1.5 kb）占5.1%，等位基因C类型（约1.2 kb）占3.4%，频率不同。在所有中，区分出19种具有I RFLP模式的等位基因亚型，鉴定出6种具有I RFLP模式的等位基因亚型。在这些样本中，分别有73个（61%）和42个（35.6%）样本通过I和I PCR-RFLP被定义为单等位基因亚型感染，而77个（65.3%）样本通过I和I RFLP的组合模式被确定为混合等位基因亚型感染。这些结果有力地表明基因具有高度多态性，特别是在从泰国-缅甸边境地区（泰国西部）采集的血样中。基因的I和I RFLP组合模式可被考虑用作泰国分离株基因分型的分子流行病学标记。
