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.4 IgG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.
Detection of GⅡ.4 Norovirus Specific IgG in Animal Serum Using A Newly Developed Luciferase Immunosorbent Assay
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.4 immunoglobulin(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 TgG in humans and different species of animals.The wide distribution of IgG antibodies against HuNoV suggests that NoV may be a zoonotic pathogen.
GⅡ.4 norovirusLuciferase immunosorbent assayIgG antibodyDomestic and wild animals
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