Siqueira Jones Anderson Monteiro, Bandeira Renato da Silva, Oliveira Darleise de Souza, Dos Santos Liann Filiphe Pereira, Gabbay Yvone Benchimol
Laboratório de Norovírus e outros Vírus Gastroentéricos-LNVE, Seção de Virologia-SAVIR, Instituto Evandro Chagas-IEC, Secretaria de Vigilância em Saúde, Ministério da Saúde, Ananindeua, Pará, Brazil.
Seção de Virologia-SAVIR, Instituto Evandro Chagas-IEC, Secretaria de Vigilância em Saúde, Ministério da Saúde, Ananindeua, Pará, Brazil.
PLoS One. 2017 Jun 12;12(6):e0178909. doi: 10.1371/journal.pone.0178909. eCollection 2017.
A chronologically comprehensive 30-year study was conducted that involved children living in Belém, in the Amazon region of Northern Brazil, who participated in eight different studies from October 1982 to April 2011. The children were followed either in the community or in health units and hospitals in order to identify the norovirus genotypes involved in infections during this time. A total of 2,520 fecal specimens were obtained and subjected to RT-PCR and nucleotide sequencing for regions A, B, C, D and P2 of the viral genome. An overall positivity of 16.9% (n = 426) was observed, and 49% of the positive samples were genotyped (208/426), evidencing the presence of several genotypes as follows: Polymerase gene (GI.P4, GII.Pa, GII.Pc, GII.Pe, GII.Pg, GII.Pj, GII.P3, GII.P4, GII.P6, GII.P7, GII.P8, GII.P12, GII.P13, GII.P14, GII.P21, GII.P22), and VP1 gene (GI.3, GI.7, GII.1, GII.2, GII.3, GII.4, GII.6, GII.7, GII.8, GII.10, GII.12, GII.14, GII.17, GII.23). The GII.P4/GII.4 genotype determined by both open reading frames (ORFs) (partial polymerase and VP1 genes) was found for 83 samples, and analyses of the subdomain P2 region showed 10 different variants: CHDC (1970s), Tokyo (1980s), Bristol_1993, US_95/96, Kaiso_2003, Asia_2003, Hunter_2004, Yerseke_2006a, Den Haag_2006b (subcluster "O") and New Orleans_2009. Recombination events were confirmed in 47.6% (n = 20) of the 42 samples with divergent genotyping by ORF1 and ORF2 and with probable different breakpoints within the viral genome. The evolutionary analyses estimated a rate of evolution of 1.02 x 10-2 and 9.05 x 10-3 subs./site/year using regions C and D from the VP1 gene, respectively. The present research shows the broad genetic diversity of the norovirus that infected children for 30 years in Belém. These findings contribute to our understanding of noroviruses molecular epidemiology and viral evolution and provide a baseline for vaccine design.
开展了一项历时30年的全面研究,涉及居住在巴西北部亚马逊地区贝伦市的儿童,这些儿童在1982年10月至2011年4月期间参与了八项不同的研究。对这些儿童在社区或卫生单位及医院进行随访,以确定这段时间内感染所涉及的诺如病毒基因型。共采集了2520份粪便标本,并对病毒基因组的A、B、C、D和P2区域进行逆转录聚合酶链反应(RT-PCR)和核苷酸测序。总体阳性率为16.9%(n = 426),49%的阳性样本进行了基因分型(208/426),证实存在以下几种基因型:聚合酶基因(GI.P4、GII.Pa、GII.Pc、GII.Pe、GII.Pg、GII.Pj、GII.P3、GII.P4、GII.P6、GII.P7、GII.P8、GII.P12、GII.P13、GII.P14、GII.P21、GII.P22)和VP1基因(GI.3、GI.7、GII.1、GII.2、GII.3、GII.4.GII.6、GII.7、GII.8、GII.10、GII.12、GII.14、GII.17、GII.23)。通过两个开放阅读框(ORF)(部分聚合酶和VP1基因)确定的GII.P4/GII.4基因型在83个样本中被发现,对P2区域亚结构域的分析显示有10种不同变体:CHDC(20世纪70年代)、东京(20世纪80年代)、布里斯托尔_1993、美国_95/96、凯索_2003、亚洲_2003、亨特_2004、耶尔塞克_2006a、海牙_2006b(亚群“O”)和新奥尔良_2009。在42个通过ORF1和ORF2基因分型不同且病毒基因组内可能存在不同断点的样本中,47.6%(n = 20)确认存在重组事件。进化分析估计,使用VP1基因的C和D区域,进化速率分别为1.02×10⁻²和9.05×10⁻³个替换/位点/年。本研究显示了在贝伦感染儿童达30年之久的诺如病毒具有广泛的遗传多样性。这些发现有助于我们了解诺如病毒的分子流行病学和病毒进化,并为疫苗设计提供基线。