Kuang Xiaozhou, Teng Zheng, Zhang Xi
Shanghai Municipal Center for Disease Control and Prevention, 1380 Zhongshan Road (west), Shanghai, 200336 China.
Gut Pathog. 2019 Jul 26;11:40. doi: 10.1186/s13099-019-0321-x. eCollection 2019.
With the help of an existing citywide comprehensive surveillance on gastroenteritis outpatients, although norovirus genogroup II (NoV GII) was tested routinely, its genotypes were never investigated systematically on a municipal level. This study aimed to understand the prevalence, major genotypes and evolutional trends of NoV GII in Shanghai during the period of 2016-2018, and to provide molecular bases for early warning for any potential NoV outbreaks.
27 sentinel hospitals from all 16 districts were recruited by stratified probability proportional to size (PPS) method in Shanghai comprehensive diarrhea surveillance programme. Stool samples were collected and screened for NoV GII by real-time reverse transcription polymerase chain reaction (qRT-PCR). For samples that were positive in qRT-PCR, conventional RT-PCR was performed to amplify the ORF1-ORF2 junction of NoV GII gene. Generated sequences were typed by RIVM online genotyping tool, and then strains of interest were analyzed phylogenetically using MEGA 6.0.
A total of 7883 stool samples were collected from diarrhea outpatients, among which 6474 were from adults and 1409 were from children. 13.66% (1077 cases) were screened positive in qRT-PCR for NoV GII, from which 71.96% (775 cases) were sequenced successfully. The top three genotypes were GII.Pe/GII.4 (37%), GII.P17/GII.17 (26%) and GII.P16/GII.2 (17%). While GII.Pe/GII.4 detection rate decreased significantly over the 3 years (from 48.4 to 20.9%); GII.P16/GII.2 appeared for the first time in October 2016 and rose rapidly to 27.0% in 2017, but fell back to 23.4% in 2018. Meanwhile there was a significant increase for both GII.P12/GII.3 and GII.P7/GII.6 recombinant genotypes detected in adult population in 2018. Phylogenic analysis revealed the existence of multiple gene clusters within both of these recombinant genotypes.
Unlike the alternating circulation of GII.4 and non-GII.4 NoV observed in 2016 or 2017, the genotype profile of NoV GII in 2018 was characterized by the co-prevalence of multiple recombinant genotypes. A recent increase in detection rate in less reported recombinant genotypes such as GII.P12/GII.3 and GII.P7/GII.6 among adult population calls for a continuing close monitoring on NoV GII genotypes in case of potential local outbreaks.
借助现有的全市范围的肠胃炎门诊患者综合监测,虽然诺如病毒II基因组(NoV GII)进行了常规检测,但从未在市级层面系统地调查过其基因型。本研究旨在了解2016 - 2018年期间上海NoV GII的流行情况、主要基因型和进化趋势,为诺如病毒潜在爆发的早期预警提供分子依据。
在上海综合腹泻监测项目中,采用按规模大小分层的概率比例抽样(PPS)方法,从16个区的27家哨点医院招募对象。收集粪便样本,通过实时逆转录聚合酶链反应(qRT-PCR)筛查NoV GII。对于qRT-PCR呈阳性的样本,进行常规RT-PCR以扩增NoV GII基因的ORF1-ORF2连接区。生成的序列通过荷兰国家公共卫生与环境研究所(RIVM)在线基因分型工具进行分型,然后使用MEGA 6.0对感兴趣的毒株进行系统发育分析。
共收集了7883份腹泻门诊患者的粪便样本,其中6474份来自成人,1409份来自儿童。13.66%(1077例)的样本经qRT-PCR筛查NoV GII呈阳性,其中71.96%(775例)成功测序。前三位基因型为GII.Pe/GII.4(37%)、GII.P17/GII.17(26%)和GII.P16/GII.2(17%)。虽然GII.Pe/GII.4的检出率在3年期间显著下降(从48.4%降至20.9%);GII.P16/GII.2于2016年10月首次出现,并在2017年迅速升至27.0%,但在2018年回落至23.4%。同时,2018年在成人中检测到的GII.P12/GII.3和GII.P7/GII.6重组基因型均显著增加。系统发育分析显示这两种重组基因型中都存在多个基因簇。
与2016年或2017年观察到的GII.4和非GII.4诺如病毒交替流行不同,2018年NoV GII的基因型特征是多种重组基因型共同流行。近期在成人中报道较少的重组基因型如GII.P12/GII.3和GII.P7/GII.6的检出率有所增加,这就要求持续密切监测NoV GII基因型,以防潜在的局部爆发。