Peralta Regina Helena Saramago, Velásquez Jorge Néstor, Cunha Flavia de Souza, Pantano María Laura, Sodré Fernando Campos, Silva Sidnei da, Astudillo Osvaldo Germán, Peralta José Mauro, Carnevale Silvana
Departamento de Patologia, Faculdade de Medicina, Universidade Federal Fluminense, Niterói, RJ, Brasil.
Hospital Municipal de Infecciosas Dr Francisco Javier Muñiz, Buenos Aires, Argentina.
Mem Inst Oswaldo Cruz. 2016 Jan;111(1):30-6. doi: 10.1590/0074-02760150303.
The identification and characterisation of Cryptosporidium genotypes and subtypes are fundamental to the study of cryptosporidiosis epidemiology, aiding in prevention and control strategies. The objective was to determine the genetic diversity of Cryptosporidium in samples obtained from hospitals of Rio de Janeiro, Brazil, and Buenos Aires, Argentina. Samples were analysed by microscopy and TaqMan polymerase chain reaction (PCR) assays for Cryptosporidium detection, genotyped by nested-PCR-restriction fragment length polymorphism (RFLP) analysis of the 18S rRNA gene and subtyped by DNA sequencing of the gp60 gene. Among the 89 samples from Rio de Janeiro, Cryptosporidium spp were detected in 26 by microscopy/TaqMan PCR. In samples from Buenos Aires,Cryptosporidium was diagnosed in 15 patients of the 132 studied. The TaqMan PCR and the nested-PCR-RFLP detected Cryptosporidium parvum, Cryptosporidium hominis, and co-infections of both species. In Brazilian samples, the subtypes IbA10G2 and IIcA5G3 were observed. The subtypes found in Argentinean samples were IbA10G2, IaA10G1R4, IaA11G1R4, and IeA11G3T3, and mixed subtypes of Ia and IIa families were detected in the co-infections. C. hominis was the species more frequently detected, and subtype family Ib was reported in both countries. Subtype diversity was higher in Buenos Aires than in Rio de Janeiro and two new subtypes were described for the first time.
隐孢子虫基因型和亚型的鉴定与特征分析是隐孢子虫病流行病学研究的基础,有助于制定预防和控制策略。目的是确定从巴西里约热内卢和阿根廷布宜诺斯艾利斯的医院采集的样本中隐孢子虫的遗传多样性。通过显微镜检查和TaqMan聚合酶链反应(PCR)检测样本中的隐孢子虫,通过对18S rRNA基因进行巢式PCR-限制性片段长度多态性(RFLP)分析进行基因分型,并通过gp60基因的DNA测序进行亚型分型。在里约热内卢的89个样本中,通过显微镜检查/TaqMan PCR在26个样本中检测到隐孢子虫属。在布宜诺斯艾利斯的样本中,在所研究的132名患者中有15名被诊断为隐孢子虫感染。TaqMan PCR和巢式PCR-RFLP检测到微小隐孢子虫、人隐孢子虫以及两种物种的混合感染。在巴西样本中,观察到亚型IbA10G2和IIcA5G3。在阿根廷样本中发现的亚型为IbA10G2、IaA10G1R4、IaA11G1R4和IeA11G3T3,在混合感染中检测到Ia和IIa家族的混合亚型。人隐孢子虫是检测到的最常见物种,两个国家均报告了亚型家族Ib。布宜诺斯艾利斯的亚型多样性高于里约热内卢,并且首次描述了两种新亚型。