Woo Patrick C Y, Lau Susanna K P, Tsoi Hoi-wah, Chan Kwok-hung, Wong Beatrice H L, Che Xiao-yan, Tam Victoria K P, Tam Sidney C F, Cheng Vincent C C, Hung Ivan F N, Wong Samson S Y, Zheng Bo-jian, Guan Yi, Yuen Kwok-yung
Department of Microbiology, University of Hong Kong, University Pathology Building, Queen Mary Hospital, Hong Kong.
Lancet. 2004 Mar 13;363(9412):841-5. doi: 10.1016/S0140-6736(04)15729-2.
Although the genome of severe acute respiratory syndrome coronavirus (SARS-CoV) has been sequenced and a possible animal reservoir identified, seroprevalence studies and mass screening for detection of subclinical and non-pneumonic infections are still lacking.
We cloned and purified the nucleocapsid protein and spike polypeptide of SARS-CoV and examined their immunogenicity with serum from patients with SARS-CoV pneumonia. An ELISA based on recombinant nucleocapsid protein for IgG detection was tested with serum from 149 healthy blood donors who donated 3 years previously and with serum positive for antibodies against SARS-CoV (by indirect immunofluorescence assay) from 106 patients with SARS-CoV pneumonia. The seroprevalence of SARS-CoV was studied with the ELISA in healthy blood donors who donated during the SARS outbreak in Hong Kong, non-pneumonic hospital inpatients, and symptom-free health-care workers. All positive samples were confirmed by two separate western-blot assays (with recombinant nucleocapsid protein and recombinant spike polypeptide).
Western-blot analysis showed that the nucleocapsid protein and spike polypeptide of SARS-CoV are highly immunogenic. The specificity of the IgG antibody test (ELISA with positive samples confirmed by the two western-blot assays) was 100%, and the sensitivity was 94.3%. Three of 400 healthy blood donors who donated during the SARS outbreak and one of 131 non-pneumonic paediatric inpatients were positive for IgG antibodies, confirmed by the two western-blot assays (total, 0.48% of our study population).
Our findings support the existence of subclinical or non-pneumonic SARS-CoV infections. Such infections are more common than SARS-CoV pneumonia in our locality.
尽管严重急性呼吸综合征冠状病毒(SARS-CoV)的基因组已被测序,且已确定了可能的动物宿主,但仍缺乏针对亚临床和非肺炎性感染检测的血清流行率研究及大规模筛查。
我们克隆并纯化了SARS-CoV的核衣壳蛋白和刺突多肽,并用SARS-CoV肺炎患者的血清检测了它们的免疫原性。基于重组核衣壳蛋白的IgG检测ELISA法,用3年前献血的149名健康献血者的血清以及106名SARS-CoV肺炎患者中抗SARS-CoV抗体呈阳性(通过间接免疫荧光法检测)的血清进行了测试。我们用ELISA法对香港SARS疫情期间献血的健康献血者、非肺炎性住院患者和无症状医护人员进行了SARS-CoV血清流行率研究。所有阳性样本均通过两种独立的蛋白质印迹分析(使用重组核衣壳蛋白和重组刺突多肽)进行了确认。
蛋白质印迹分析表明,SARS-CoV的核衣壳蛋白和刺突多肽具有高度免疫原性。IgG抗体检测(通过两种蛋白质印迹分析确认阳性样本的ELISA法)的特异性为100%,敏感性为94.3%。在SARS疫情期间献血的400名健康献血者中有3名以及131名非肺炎性儿科住院患者中有1名IgG抗体呈阳性,经两种蛋白质印迹分析确认(占我们研究人群的0.48%)。
我们的研究结果支持亚临床或非肺炎性SARS-CoV感染的存在。在我们当地,此类感染比SARS-CoV肺炎更为常见。