Department of Anatomy and Biology, College of Medicine, University of Wasit, Wasit, Iraq.
Department of Microbiology, College of Medicine, University of Wasit, Wasit, Iraq.
Arch Razi Inst. 2023 Feb 28;78(1):337-343. doi: 10.22092/ARI.2022.358947.2336. eCollection 2023 Feb.
Amebiasis is caused by , a protozoan that is found worldwide. The degree of pathogenesis of clinical isolates varies greatly. This study was aimed to molecular identification of in children using the nested polymerase chain reaction (nPCR), and then, a genotyping of positive isolates using the quantitative PCR (qPCR) assay through targeting serine-rich protein (SREHP) gene. For this purpose, a total of 50 bloody diarrheic stool samples were collected from the children attended to the Al-Zahraa' Teaching Hospital and Alkut Hospital for Gynecology, Obstetric and Pediatrics (Alkut, Wasit, Iraq) were subjected to the present study from September to December 2021. Firstly, the extracted DNAs that amplified using specific primers through targeting gene and tested using nPCR assay were revealed an overall 48% (24/50) positive samples for . For genotyping, our results were detected an existence of four different genotypes (I, II, III and IV) with a significant prevalence of Genotype-II (54.17%) when compared to Genotype-I (20.83%), Genotype-III (12.5%) and Genotype- IV (12.5%). In addition, results of melting temperature of targeted genotypes were 84ºC, 83 - 83.5ºC, 82.5ºC and 81ºC for Genotype-I, II, III and IV, respectively. In conclusion, molecular amplification of gene was revealed the large prevalence of among bloody diarrheic children of study areas; while, amplification of SREHP gene was reflected the widespread phenotypic variation of the Genotype-II suggesting the high ability of this genotype to spread infection in children. In different endemic areas as Iraq, the utilization of high-resolution genotyping methods showed the extremely polymorphic genetic structure of this parasite.
溶组织内阿米巴病是由 引起的，一种在世界范围内都有发现的原生动物。临床分离株的发病机制程度差异很大。本研究旨在使用巢式聚合酶链反应（nPCR）对儿童中的 进行分子鉴定，然后使用针对富含丝氨酸的蛋白（SREHP）基因的定量 PCR（qPCR）检测对阳性 分离株进行基因分型。为此，本研究共收集了 2021 年 9 月至 12 月期间在 Al-Zahraa' 教学医院和 Alkut 妇产科医院（Wasit，伊拉克）就诊的 50 例血性腹泻粪便样本。首先，通过针对 基因使用特异性引物进行提取的 DNA 通过 nPCR 检测呈阳性的样本占 48%（24/50）。对于基因分型，我们的结果检测到存在四种不同的基因型（I、II、III 和 IV），与基因型-I（20.83%）相比，基因型-II（54.17%）显著更为流行，基因型-III（12.5%）和基因型-IV（12.5%）。此外，目标基因型的熔点温度结果分别为 84°C、83-83.5°C、82.5°C 和 81°C，基因型-I、II、III 和 IV。总之，基因的分子扩增揭示了研究地区血性腹泻儿童中 的高流行率；而 SREHP 基因的扩增反映了基因型-II 的表型变异广泛，表明该基因型在儿童中传播感染的能力很强。在像伊拉克这样的不同流行地区，利用高分辨率基因分型方法显示了该寄生虫具有极高的遗传结构多态性。
