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[腹泻病例中芽囊原虫的遗传多样性:亚型和等位基因的鉴定]

[Genetic Diversity of Blastocystis in Diarrheal Cases: Identification of Subtypes and Alleles].

作者信息

Tileklioğlu Evren, Ertabaklar Hatice

机构信息

Aydın Adnan Menderes University Faculty of Medicine, Department of Parasitology, Aydın, Türkiye.

出版信息

Mikrobiyol Bul. 2024 Apr;58(2):196-208. doi: 10.5578/mb.202498207.

Abstract

Blastocystis spp. are the most common intestinal protozoan parasites detected in human stool samples. While identified long before today, its pathogenicity remains controversial. It is generally asymptomatic but in symptomatic cases, many gastrointestinal symptoms, especially diarrhea, have been associated with Blastocystis infection. In recent years, the relationship between the symptoms observed in cases and Blastocystis subtypes (ST) has been reported. The aim of this study was to detect Blastocystis in diarrheal cases admitted to the Aydın Adnan Menderes University Faculty of Medicine, Department of Parasitology Laboratory, to determine subtypes and allele diversity and to investigate its relationship with clinical symptoms. For this purpose, diarrheal stool samples of 200 cases were included in the study and their demographic characteristics (age, gender, residence) and clinical findings (abdominal pain, dyspepsia, nausea-vomiting, weakness, weight loss, anal itching, rash, urticaria) were recorded. Blastocystis was detected by direct microscope method (DM) and by molecular analyses which were performed with polymerase chain reaction (PCR). Subtype diversity was determined based on DNA sequence analysis by PCR targeting the Blastocystis ribosomal ribonucleic acid small subunit (SSU rRNA) gene. In addition, alleles related to Blastocystis subtypes were determined and statistically compared between all data and clinical findings. In the current study, Blastocystis was detected in 31 (15.5%) samples by DM and in 35 (17.5%) samples by PCR specific to the Blastocystis SSU rRNA gene among 200 diarrheal stool samples. No statistical difference was detected between Blastocystis and demographic characteristics. Dyspepsia and nausea-vomiting symptoms differed significantly in cases with Blastocystis compared to negative ones (p= 0.0025, p= 0.0498). Blastocystis subtype was detected in 33 samples by SSU rRNA sequence analysis, and the subtype distribution was ST1 (n= 10, 30.3%), ST2 (n= 4, 12.1%) and ST3 (n= 19, 57.6%). In the statistical evaluation between clinical findings and Blastocystis subtypes, a relationship was found between dyspepsia and Blastocystis ST3 (p= 0.0039). The allele diversity of Blastocystis subtypes was determined as allele 4 (10/10) in all ST1, allele 11 (2/4) and 12 (2/4) in ST2, allele 34 (14/19), 36 (4/19), and 38 (1/19) in ST3. In conclusion, our study provides important data on the molecular epidemiological characteristics of the Blastocystis by determining positivity, subtypes and alleles in diarrheal cases. Therefore, within the scope of the one health approach, comprehensive molecular epidemiological studies are required to determine the presence and genotypes of Blastocystis in human, animal and environmental samples.

摘要

芽囊原虫是在人类粪便样本中检测到的最常见的肠道原生动物寄生虫。虽然早在今日之前就已被发现,但其致病性仍存在争议。它通常无症状,但在有症状的病例中,许多胃肠道症状,尤其是腹泻,已被认为与芽囊原虫感染有关。近年来,已报道了病例中观察到的症状与芽囊原虫亚型(ST)之间的关系。本研究的目的是在艾登阿德南·曼德雷斯大学医学院寄生虫学实验室收治的腹泻病例中检测芽囊原虫,确定亚型和等位基因多样性,并研究其与临床症状的关系。为此,本研究纳入了200例腹泻粪便样本,并记录了其人口统计学特征(年龄、性别、居住地)和临床发现(腹痛、消化不良、恶心呕吐、虚弱、体重减轻、肛门瘙痒、皮疹、荨麻疹)。通过直接显微镜检查法(DM)和聚合酶链反应(PCR)进行的分子分析检测芽囊原虫。基于针对芽囊原虫核糖体核糖核酸小亚基(SSU rRNA)基因的PCR的DNA序列分析确定亚型多样性。此外,确定了与芽囊原虫亚型相关的等位基因,并在所有数据和临床发现之间进行了统计学比较。在本研究中,在200份腹泻粪便样本中,通过DM在31份(15.5%)样本中检测到芽囊原虫,通过针对芽囊原虫SSU rRNA基因的PCR在35份(17.5%)样本中检测到芽囊原虫。在芽囊原虫与人口统计学特征之间未检测到统计学差异。与阴性病例相比,有芽囊原虫的病例中的消化不良和恶心呕吐症状有显著差异(p = 0.0025,p = 0.0498)。通过SSU rRNA序列分析在33份样本中检测到芽囊原虫亚型,亚型分布为ST1(n = 10,30.3%)、ST2(n = 4,12.1%)和ST3(n = 19,57.6%)。在临床发现与芽囊原虫亚型之间的统计学评估中,发现消化不良与芽囊原虫ST3之间存在关联(p = 0.0039)。芽囊原虫亚型的等位基因多样性确定为所有ST1中的等位基因4(1_0/10)、ST2中的等位基因11(2/4)和12(2/4)、ST3中的等位基因34(14/19)、36(4/19)和38(1/19)。总之,我们的研究通过确定腹泻病例中的阳性率、亚型和等位基因,提供了关于芽囊原虫分子流行病学特征的重要数据。因此,在“同一健康”方法的范围内,需要进行全面的分子流行病学研究,以确定人类、动物和环境样本中芽囊原虫的存在和基因型。

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