Association of Toxoplasma Gondii Infection With Diabetes Mellitus Using Nested-PCR and Sequencing

Sadegh Shamsinia, A. Dalimi, M. Pirestani, H. Majidiani
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Abstract

Background: Toxoplasmosis is a global public health concern with severe complications, particularly in pregnant women and immunosuppressed patients. Herein, we revealed the first genotypic evidence of Toxoplasma gondii (T. gondii) in diabetic and non-diabetic individuals referred to Imam Sajjad Hospital, Tehran, Iran, using nested-Polymerase Chain Reaction (PCR) and sequencing. Materials and Methods: We collected 98, 95, and 94 blood samples from Type-1 Diabetes Mellitus (T1DM), Type-2 Diabetes Mellitus (T2DM), and control subjects from February to August 2018, and their DNA was extracted. After amplification and visualization of dense granule antigen 6 gene (344 bp) using external and internal primer pairs, the positive samples were sent for sequencing. Results: Among 287 individuals, three T1DM patients (two IgG, one IgM) and one T2DM patient (IgG) were found positive by molecular method, which proved to be type III strain by sequencing. Conclusion: To the best of our knowledge, this is the first genotyping investigation on T. gondii in diabetic individuals. Further research should be done to better realize the association between Toxoplasma genotypes and the outcome of diabetes mellitus.
应用巢式PCR和测序技术检测弓形虫感染与糖尿病的关系
背景:弓形虫病是一个全球性的公共卫生问题,具有严重的并发症,尤其是在孕妇和免疫抑制患者中。在此,我们使用嵌套聚合酶链式反应(PCR)和测序,在转诊至伊朗德黑兰Imam Sajjad医院的糖尿病和非糖尿病患者中首次发现了弓形虫(T.gondii)的基因型证据。材料和方法:我们收集了2018年2月至8月1型糖尿病(T1DM)、2型糖尿病(T2DM)和对照受试者的98、95和94份血液样本,并提取了他们的DNA。在使用外部和内部引物对致密颗粒抗原6基因(344bp)进行扩增和可视化后,将阳性样本送去测序。结果:在287例患者中,3例T1DM患者(2例IgG,1例IgM)和1例T2DM患者(IgG)经分子方法检测呈阳性,测序证实为Ⅲ型毒株。结论:据我们所知,这是首次对糖尿病患者弓形虫进行基因分型研究。应进一步研究弓形虫基因型与糖尿病预后之间的关系。
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