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目的了解南宁市朝阳溪污水中诺如病毒(Norovirus,NV)污染状况及其基因分布。方法应用实时荧光逆转录-聚合酶链(Realtime RT-PCR)方法对2011年采集的72份朝阳溪污水进行NV检测,阳性标本进行基因克隆和序列测定,并进行亲缘关系分析。结果 72份污水均能检出NV,基因Ⅰ组(GenogroupⅠ,GⅠ)NV、基因Ⅱ组(GenogroupⅡ,GⅡ)NV阳性率分别为70.83%和100.00%。经亲缘关系树分析发现,测序成功的239株阳性克隆,共包含有16种不同亚型的NV,GⅠ(GⅠ.1、GⅠ.3、GⅠ.4、GⅠ.6、GⅠ.9、GⅠ.10、GⅠ.13、GⅠ.14、)、GⅡ(GⅡ.1、GⅡ.2、GⅡ.3、GⅡ.4、GⅡ.5、GⅡ.6、GⅡ.12、GⅡ.14)各8种。污水中GⅠ和GⅡNV代表株与广西急性腹泻病例的各亚型代表株的亲缘关系密切。在污水中还检测到37株在广西腹泻病例中未检测到的基因亚型NV,占总NV检出株数的15.48%(37/239),其中GⅠ有6个亚型、GⅡ有2个亚型。NV各亚型中检出比例最高的是GⅡ.14,占31.80%(76/239)、GⅠ.4次之,占18.83%(45/239)。在污水中同时检出GⅡ.4 2006年变异株与2010年变异株。结论污水监测能从侧面反映有多个基因亚型NV在南宁市人群中流行,但一些基因亚型的NV可能以隐性感染为主。
Objective To investigate the status and gene distribution of Norovirus (NV) in Chaoyangxi wastewater in Nanning City. Methods 72 samples of Chaoyangxi wastewater collected in 2011 were detected by Realtime RT-PCR. The positive samples were cloned and sequenced, and the genetic relationship was analyzed. Results Genotype 2 (Genogroup Ⅰ, GⅠ) NV and Genogroup Ⅱ (GⅡ) NV were detected in 72 sepsis samples. The positive rates of NV were 70.83% and 100.00%, respectively. The phylogenetic tree analysis showed that there were 239 positive clones successfully sequenced and contained 16 different subtypes of NV, GⅠ (GⅠ.1, GⅠ.3, GⅠ.4, GⅠ.6, GⅠ.9, GⅠ. 10, GⅠ.13, GⅠ.14), GⅡ (GⅡ.1, GⅡ.2, GⅡ.3, GⅡ.4, GⅡ.5, GⅡ.6, GⅡ.12, GⅡ.14). The representative strains of GⅠ and GⅡNV in sewage were closely related to the representatives of each subtype of acute diarrhea cases in Guangxi. 37 subtypes of genotypes not detected in Guangxi diarrhea cases were also detected in sewage samples, accounting for 15.48% (37/239) of the total number of strains detected in the genus. Among them, there were 6 subtypes of GⅠ and 2 subtypes of GⅡ type. The highest proportion of the detected subtypes of NV was GII.14, accounting for 31.80% (76/239), followed by GⅠ.4, accounting for 18.83% (45/239). In the sewage simultaneously detected G Ⅱ.4 2006 variant and 2010 variant. Conclusion Sewage monitoring can reflect multiple genotypes of subtype NV in the population of Nanning city from the side, but NV of some gene subtypes may be dominated by latent infection.