Monoclonal antibody-based antigen capture enzyme-linked immunosorbent assay reveals high sensitivity of the nucleocapsid protein in acute-phase sera of severe acute respiratory syndrome patients
Monoclonal antibody-based antigen capture enzyme-linked immunosorbent assay reveals high sensitivity of the nucleocapsid protein in acute-phase sera of severe acute respiratory syndrome patients
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DOI：
10.1128/cdli.12.1.135-140.2005
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发表时间：
2005-01-01
期刊：
影响因子：
--
通讯作者：
Che, XY
中科院分区：
文献类型：
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作者：
Di, B;Hao, W;Che, XY
准确及时地诊断严重急性呼吸综合征冠状病毒（SARS-CoV）感染是防止全球再次爆发的关键一步。在这项研究中，从最初报告感染 SARS-CoV 的 643 名患者收集了 829 份血清标本。使用基于针对 SARS-CoV N 蛋白的单克隆抗体的抗原捕获酶联免疫吸附测定 (ELISA) 检测血清中 SARS-CoV 的 N 蛋白，并与来自健康供体和非 SARS 发热患者的 197 份对照血清样本进行比较。 N蛋白检测分析结果与血清学分析数据直接相关。中和试验呈阳性的 27 名 SARS 患者中，从症状出现后 1 天到 10 天收集的 24 份血清中，100% N 蛋白呈阳性。在该组中第 11 天之后未检测到 N 蛋白。 298例血清学确诊患者的414份样本中，发病后1～5、6～10、11～15、16～20天采集的血清N蛋白阳性率分别为92.9%、69.8%、36.4%和21.1%。对于来自 248 名血清学检测阴性患者的 294 份血清，比率分别为 25.6%、16.7%、9.3% 和 0%。在 66 名最初被怀疑为 SARS 病例但血清学证明为 SARS 阴性的患者以及来自健康捐献者和非 SARS 发热患者的 197 份血清样本中未检测到 N 蛋白。测定的特异性为100%。此外，在广州SARS复发期间从4名患者采集的16份血清中，在症状出现后7至9天采集的5份血清中N蛋白呈阳性。 27名中和试验阳性SARS患者和298名血清学确诊患者在症状出现后第3天至第5天期间，N蛋白检测显示出较高的阳性率，为96%至100%。从第10天开始，N蛋白检出率持续下降，症状出现后第19天后，N蛋白检出率持续下降。总之，抗原捕获ELISA显示SARS患者急性期血清中N蛋白检出率较高，这有助于SARS的早期诊断。
Accurate and timely diagnosis of severe acute respiratory syndrome coronavirus (SARS-CoV) infection is a critical step in preventing another global outbreak. In this study, 829 serum specimens were collected from 643 patients initially reported to be infected with SARS-CoV. The sera were tested for the N protein of SARS-CoV by using an antigen capture enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibodies against the N protein of SARS-CoV and compared to 197 control serum samples from healthy donors and non-SARS febrile patients. The results of the N protein detection analysis were directly related to the serological analysis data. From 27 SARS patients who tested positive with the neutralization test, 100% of the 24 sera collected from I to 10 days after the onset of symptoms were positive for the N protein. N protein was not detected beyond day 11 in this group. The positive rates of N protein for sera collected at I to 5, 6 to 10, 11 to 15, and 16 to 20 days after the onset of symptoms for 414 samples from 298 serologically confirmed patients were 92.9, 69.8, 36.4, and 21.1%, respectively. For 294 sera from 248 serological test-negative patients, the rates were 25.6, 16.7, 9.3, and 0%, respectively. The N protein was not detected in 66 patients with cases of what was initially suspected to be SARS but serologically proven to be negative for SARS and in 197 serum samples from healthy donors and non-SARS febrile patients. The specificity of the assay was 100%. Furthermore, of 16 sera collected from four patients during the SARS recurrence in Guangzhou, 5 sera collected from 7 to 9 days after the onset of symptoms were positive for the N protein. N protein detection exhibited a high positive rate, 96 to 100%, between day 3 and day 5 after the onset of symptoms for 27 neutralization test-positive SARS patients and 298 serologically confirmed patients. The N protein detection rate continually decreased beginning with day 10, and N protein was not detected beyond day 19 after the onset of symptoms. In conclusion, an antigen capture ELISA reveals a high N protein detection rate in acute-phase sera of patients with SARS, which makes it useful for early diagnosis of SARS.