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诺如病毒（Norovirus, NoV）是引起全球范围内人类急性胃肠炎（Acutegastroenteritis, AGE）的主要病原体，对全球健康和经济造成了重大威胁。已有研究在不同种类的动物粪便中检出人类诺如病毒（Human norovirus,HuNoV）的核酸或在家畜血清中检测出可识别HuNoV病毒样颗粒（Virus like particles,VLP）的抗体，这引起了公众对NoV是否为潜在人兽共患病的关注与讨论。本研究基于HuNoV主要流行株GⅡ.4建立了一种检测GⅡ.4IgG的荧光素酶免疫吸附试验（Luciferase immunosorbent assay,LISA）。LISA法具有较高的灵敏度（94.87%）和特异度（98.18%）；最低检测限是ELISA法的2～16倍；GⅡ.4-LISA检测人血清的批间和批内变异系数值均小于10%，重复性良好。GⅡ.4 IgG在狗、猪、蝙蝠、大鼠和鼩鼱等不同的家畜和野生动物中均有检出，阳性率在3.37%～39.53%之间，其中蝙蝠的检出率最高（39.53%），狗次之（9.72%）。综上所述，本研究建立的GⅡ.4-LISA方法适用于人和不同种属动物中GⅡ.4特异性IgG检测。针对HuNoV的IgG抗体广泛分布，提示NoV可能是潜在的人畜共患病。
Abstract：Norovirus(NoV) is the leading cause of acute gastroenteritis. It poses a major threat to the health and economy of societies worldwide. The nucleic acids of human norovirus(HuNoV) have been detected in the feces of different species of animals. Antibodies to HuNoV virus-like particles have been detected in the serum of domestic animals. These phenomena have attracted public attention and discussion on whether NoV is potential zoonotic pathogen. We developed a new luciferase immunosorbent assay(LISA) to detect GⅡ. 4immunoglobulin(Ig)G based on the main epidemic strain of HuNoV GⅡ. 4. Our LISA had high sensitivity(94.87%) and specificity(98.18%). The limit of detection was 2 – 16-times higher than that of an enzyme-linked immunosorbent assay. The inter-and intra-assay coefficients of variation of GⅡ.4-LISA were <10%, indicating good repeatability. GⅡ.4 IgG was detected in different domestic and wild animals(dogs, pigs, bats, rats and shrews) with positive rates ranging from 3.37% to 39.53%. The positive rate for the bat was the highest(39.53%), followed by the dog(9.72%). In conclusion, the GⅡ.4-LISA established in this study was suitable for the detection of GⅡ.4-specific IgG in humans and different species of animals. The wide distribution of IgG antibodies against HuNoV suggests that NoV may be a zoonotic pathogen.
[1] Thorne LG, Goodfellow IG. Norovirus gene expression and replication[J]. J Gen Virol, 2014, 95(pt 2):278-291. DOI:10. 1099/vir. 0. 059634-0.
[2] VinjéJ. Advances in laboratory methods for detection and typing of norovirus[J]. J Clin Microbiol, 2015, 53(2):373-381. DOI:10. 1128/jcm. 01535-14.
[3] Wang QH, Han MG, Cheetham S, et al. Porcine noroviruses related to human noroviruses[J]. Emerg Infect Dis, 2005, 11(12):1874-1881. DOI:10. 3201/eid1112. 050485.
[4] Chhabra P, de Graaf M, Parra GI, et al. Updated classification of norovirus genogroups and genotypes[J].J Gen Virol, 2019, 100(10):1393-1406. DOI:10. 1099/jgv. 0. 001318.
[5] de Graaf M, van Beek J, Koopmans MPG. Human norovirus transmission and evolution in a changing world[J]. Nat Rev Microbiol, 2016, 14:421-433. DOI:10. 1038/nrmicro. 2016. 48.
[6] Yoon HJ, Hur SH. Determination of safe contrast media dosage to estimated glomerular filtration rate ratios to avoid contrast-induced nephropathy after elective percutaneous coronary intervention[J]. Korean Circ J, 2011, 41(5):265-271. DOI:10. 4070/kcj. 2011. 41. 5. 265.
[7] Ramani S, Atmar RL, Estes MK. Epidemiology of human noroviruses and updates on vaccine development[J]. Curr Opin Gastroenterol, 2014, 30(1):25-33.DOI:10. 1097/mog. 0000000000000022.
[8] Patel MM, Widdowson MA, Glass RI, et al.Systematic literature review of role of noroviruses in sporadic gastroenteritis[J]. Emerg Infect Dis, 2008, 14(8):1224-1231. DOI:10. 3201/eid1408. 071114.
[9] Netzler NE, Enosi Tuipulotu D, White PA. Norovirus antivirals:Where are we now?[J]. Med Res Rev,2019, 39(3):860-886. DOI:10. 1002/med. 21545.
[10]Widdowson MA, Rockx B, Schepp R, et al. Detection of serum antibodies to bovine norovirus in veterinarians and the general population in the Netherlands[J]. J Med Virol, 2005, 76(1):119-128. DOI:10. 1002/jmv. 20333.
[11]Mesquita JR, Costantini VP, Cannon JL, et al.Presence of antibodies against genogroup VI norovirus in humans[J]. Virol J, 2013, 10:176. DOI:10. 1186/1743-422x-10-176.
[12]Humphrey TJ, Cruickshank JG, Cubitt WD. An outbreak of calicivirus associated gastroenteritis in an elderly persons home. A possible zoonosis?[J]. J Hyg(Lond), 1984, 93(2):293-299. DOI:10. 1017/s0022172400064822.
[13]Summa M, von Bonsdorff CH, Maunula L. Pet dogs—a transmission route for human noroviruses?[J]. J Clin Virol, 2012, 53(3):244-247. DOI:10. 1016/j.jcv. 2011. 12. 014.
[14]Caddy SL, de Rougemont A, Emmott E, et al.Evidence for human norovirus infection of dogs in the United Kingdom[J]. J Clin Microbiol, 2015, 53(6):1873-1883. DOI:10. 1128/jcm. 02778-14.
[15]Nakamura K, Saga Y, Iwai M, et al. Frequent detection of noroviruses and sapoviruses in swine and high genetic diversity of porcine sapovirus in Japan during Fiscal Year 2008[J]. J Clin Microbiol, 2010, 48(4):1215-1222. DOI:10. 1128/jcm. 02130-09.
[16]Bucardo F, González F, Reyes Y, et al.Seroprevalence in household raised pigs indicate high exposure to GII noroviruses in rural Nicaragua[J].Zoonoses Public Health, 2016, 63(8):600-607. DOI:10. 1111/zph. 12269.
[17]Liang Z, Zhang M, Wang Y, et al. Development and evaluation of a new luciferase immunosorbent assay to detect GII. 6 norovirus-specific IgG in different domestic and wild animals[J]. Front Microbiol, 2023, 14:1213007. DOI:10. 3389/fmicb. 2023. 1213007.
[18]Wang H, Cai Q, Liang Y, et al. A simple and highthroughput luciferase immunosorbent assay for both qualitative and semi-quantitative detection of anti-HIV-1antibodies[J]. Virus Res, 2019, 263:9-15. DOI:10. 1016/j. virusres. 2018. 12. 017.
[19]Bank-Wolf BR, K?nig M, Thiel HJ. Zoonotic aspects of infections with noroviruses and sapoviruses[J]. Vet Microbiol, 2010, 140(3-4):204-212. DOI:10. 1016/j. vetmic. 2009. 08. 021.
[20]王天禹，芜为，邓瑶，等.基于寨卡病毒NS2B蛋白特异抗原多肽的荧光素酶免疫吸附法检测寨卡病毒抗体[J].病毒学报，2019, 35(3):396-401. DOI:10. 13242/j. cnki. bingduxuebao. 003546.
[21]Hall MP, Unch J, Binkowski BF, et al. Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate[J]. ACS Chem Biol,2012, 7(11):1848-1857. DOI:10. 1021/cb3002478.
[22]Baeshen MN, Al-Hejin AM, Bora RS, et al.Production of biopharmaceuticals in E. coli:current scenario and future perspectives[J]. J Microbiol Biotechnol, 2015, 25(7):953-962. DOI:10. 4014/jmb. 1412. 12079.
[23]Wolf S, Reetz J, Johne R, et al. The simultaneous occurrence of human norovirus and hepatitis E virus in a Norway rat(Rattus norvegicus)[J]. Arch Virol, 2013,158(7):1575-1578. DOI:10. 1007/s00705-013-1646-2.
[24]Kocher JF, Lindesmith LC, Debbink K, et al. Bat caliciviruses and human noroviruses are antigenically similar and have overlapping histo-blood group antigen binding profiles[J]. mBio, 2018, 9(3):e00869-e00818. DOI:10. 1128/mbio. 00869-18.
[25]Wu Z, Yang L, Ren X, et al. Deciphering the bat virome catalog to better understand the ecological diversity of bat viruses and the bat origin of emerging infectious diseases[J]. ISME J, 2016, 10(3):609-620. DOI:10. 1038/ismej. 2015. 138.
基本信息:
DOI：10.13242/j.cnki.bingduxuebao.004582
中图分类号:R512.5;R446.6
引用信息:
[1]王宇,张敏怡,徐嘉怡等.诺如病毒GⅡ.4 IgG抗体的荧光素酶免疫吸附方法的建立及在动物血清中的应用[J].病毒学报,2024,40(05):978-985.DOI:10.13242/j.cnki.bingduxuebao.004582.
基金信息:
广东省重点领域研发项目（项目号：2022B1111020002）,题目：华南地区主要流行人类病毒模式小鼠资源库的建立及评估应用; 国家自然科学基金（项目号：31771007）,题目：GII.4型诺如病毒全人广谱中和单抗及其保守表位研究; 广东省自然科学基金（项目号：2019A1515010951）,题目：基于自然人群队列诺如病毒基因型更迭的群体免疫机制研究~~