Laboratório de Virologia Comparada e Ambiental, Instituto Oswaldo Cruz/Fiocruz, Rio de Janeiro, Rio de Janeiro, Brazil.
PLoS One. 2012;7(3):e33754. doi: 10.1371/journal.pone.0033754. Epub 2012 Mar 20.
This 15-year study aimed to determine the role of the main viruses responsible for acute infantile gastroenteritis cases in a day care center in the city of Rio de Janeiro, Brazil. From 1994 to 2008, 539 fecal samples were obtained from 23 outbreaks as well as sporadic cases that occurred in this period. The detection of Rotavirus group A (RVA), norovirus (NoV) and astrovirus (AstV) was investigated both by classical and molecular methods of viral detection. RVA was detected by enzymatic immune assay and/or polyacrylamide gel electrophoresis and genotyped by using semi-nested multiplex PCR. NoV and AstV were subsequently tested by real time PCR in all RVA-negative samples and genotyped throughout genome sequencing. Three protocols for molecular characterization of NoV nucleotide sequencing were performed with the partial nucleotide sequencing of genomic regions known as region B (polymerase gen), C and D (capsid gen).Viruses were identified in 47.7% (257/539) of the cases, and the detection rates of RVA, NoV and AstV in16.1% (87/539), 33.4% (151/452), and 6.3% (19/301), respectively. Most gastroenteritis cases were reported in autumn and winter, although NoV presented a broader monthly distribution. Viruses' detection rates were significantly higher among children aged less than 24 months old, although NoV cases were detected in all age groups. RVA genotypes as G1P[8], G9P[8], G2P[4], G3P[8] and G1+G3P[8] and RVA was no longer detected after 2005. NoV characterization revealed genotypes variability circulating in the period as GI.2, GI.3, GI.8 GII.2, GII.3, GII.4, GII.4 variants 2001 and 2006b, GII.6, GII.7, GII.12 and GII.17. AstV genotypes 1, 2, 4 and 5 were also characterized. Those data demonstrate the impact of NoV infection in cases of infantile gastroenteritis, surpassing RVA infection responsible for high morbidity rate in children under five years old.
这项为期 15 年的研究旨在确定导致巴西里约热内卢市日托中心急性婴儿肠胃炎病例的主要病毒的作用。1994 年至 2008 年,从这一时期的 23 次暴发和散发病例中获得了 539 份粪便样本。通过经典和病毒检测的分子方法检测轮状病毒 A 组(RVA)、诺如病毒(NoV)和星状病毒(AstV)。通过酶免疫测定和/或聚丙烯酰胺凝胶电泳检测到 RVA,并通过半嵌套多重 PCR 进行基因分型。在所有 RVA 阴性样本中均通过实时 PCR 检测到 NoV 和 AstV,并通过全基因组测序进行基因分型。用部分核苷酸测序法对 NoV 核苷酸测序的三个方案进行了 3 次分子特征描述,所用的基因组区域为 B 区(聚合酶基因)、C 区和 D 区(衣壳基因)。在 539 例病例中,有 47.7%(257/539)检测到病毒,RVA、NoV 和 AstV 的检出率分别为 16.1%(87/539)、33.4%(151/452)和 6.3%(19/301)。肠胃炎病例主要发生在秋季和冬季,但 NoV 的分布范围更广。尽管在所有年龄组均检测到 NoV 病例,但病毒检出率在年龄小于 24 个月的儿童中明显更高。RVA 基因型为 G1P[8]、G9P[8]、G2P[4]、G3P[8]和 G1+G3P[8],自 2005 年后不再检出 RVA。NoV 特征分析显示,在该时期流行的基因型为 GI.2、GI.3、GI.8、GII.2、GII.3、GII.4、GII.4 变异体 2001 和 2006b、GII.6、GII.7、GII.12 和 GII.17。还对 AstV 基因型 1、2、4 和 5 进行了特征描述。这些数据表明,NoV 感染对婴儿肠胃炎病例的影响超过了导致五岁以下儿童高发病率的 RVA 感染。