20例SARS患者特异性抗体变化规律

目的：了解严重急性呼吸道综合征(SARS)特异性IgM和IgG抗体的变化规律。方法：采用间接酶联免疫吸附试验(ELISA)检测20例SARS患者系列血清中特异性IgM和IgG抗体，系列血清包括患者发病后1周，2周，3周，4周，8周，12周所采集的样本。结果：20例患者发病后第1周IgM和IgM抗体均为阴性；第2周时16例IgM抗体阳性，17例IgM抗体阳性；第3周后所有患者IgG抗体阳性并持续至第12周，而IgM抗体阳性患者逐渐减少，至第12周时所有患者均为阴性。结论：SARS特异性IgG抗体消失较早，其存在是近期感染的标志；IgG抗体的持续存在可能是获得病后免疫力的标志。     

SARS患者康复期血清特异性抗体效价检测

目的：SARS患者康复期血清特异性抗体的效价。方法：采用间接酶联免疫法和双抗原夹心法。结果：SARS患者康复期血清特异性抗体IgG水平在住院第5周达最高值，IgM水平在住院第3周达到最高值。SARS患者康复期血清IgG水平是IgM的1～7倍。住院5～7周的SARS康复期患者，其血清特异性抗体检测阳性率达100％。结论：SAPS患者康复期血清特异性抗体水平在住院第5周达到最高值，其阳性检出率达100％。     

传播链和非传播链SARS患者中SARS冠状病毒抗体的分布及变化规律

目的　探讨人体对SARS CoV血清抗体反应及分布规律。方法　对 30 1例临床诊断病例的血清标本 (其中传播链上的病例 15 8例 ,非传播链病例 14 3例 ) ;采用北京华大GBI生物技术有限公司生产的间接ELISA试剂 ,进行SARS CoVIgG抗体的检测 ,部分病例还进行了SARS CoVIgM的检测。结果　传播链涉及 15条 ,最多涉及 4代 ,各条链的SARSIgG抗体阳性率为 85 70 %～ 10 0 .0 0 % ,总阳性率为 94 30 % (14 9 15 8)。SARS病例数随代数增加而减少。但各代间IgG阳性率的差异无显著意义 ;χ2 =5 11,P >0 0 5。非传播链病例的血清抗体阳性率为 12 5 9% (18 14 3)。与传播链的阳性率比较 ,两者间的差异有显著意义 ;χ2 =199 6 4 ,P <0 0 0 1。在发病后的 0～ 7d、8～ 14d、15～ 2 1d、2 2～ 30d内 ,SARS CoVIgG的累计阳性率分别为 16 6 7%、4 0 0 0 %、70 0 0 %、93 10 % ;然后进入平台期 ,并可以维持 6个月以上。在发病 7d内 ,SARS CoVIgM的累计阳性率为 16 6 7% ,8～ 14d时 ,已有 5 5 17%的标本IgM抗体阳转 ,在 2 1～ 30d时 ,抗体阳性率达高峰 (86 96 % ) ,以后迅速下降。结论　SARS感染后 ,94 %的感染者可产生IgG抗体。检测该抗体可以作为临床核实诊断的依据。SARS CoVIgG抗体在 7d时可能检出 ,一般在 4周达     

SARS病毒抗体的检测及其应用

为了解SARS患者血清抗体产生的规律 ,我们用间接免疫荧光法 (IFA)和酶联免疫吸附试验 (ELISA)对广州市第八人民医院的 4 3例住院治疗的SARS患者和 10例正常人的双份血清进行SARS抗体检测 ,同时比较了IFA和ELISA法的特点。10例正常人双份血清的SARS病毒IgM和IgG抗体均阴性。 4 3例SARS患者 ,其中 34例检出IgM占 79.1%。 37例检出IgG占 86 .10 %。SARS病毒IgG抗体的阳性率高于IgM。结果显示 ,发病的同时机体便可检测到SARS抗体IgM最迟离起病时间最长 (33d)的血清也能检测到 ,但也有患者发病第 9、2 3及 30天的第二份…     

SARS-CoV核衣壳蛋白用于SARS血清学诊断的研究

目的　研究SARS患者血清中抗SARS冠状病毒 (SARS CoV)核衣壳 (N)蛋白特异性抗体的产生规律 ,评价N蛋白在SARS诊断中的作用。方法　采用间接ELISA法测定 2 5 0例临床确诊、1 88例临床疑似、6 2例临床排除SARS患者血清中针对N蛋白IgG抗体 ,并与其SARS CoVIgG总抗体水平进行比较。结果　临床确诊病例针对N蛋白和全病毒特异性抗体IgG的阳性率均随着病程的延长而增高。N IgG在发病第 1周检出阳性率为 3.4 5 %(4 1 1 6 ) ,第 2周为 6 1 .76 %(42 6 8) ,第 3周为81 .82 %(1 8 2 2 ) ,2 2～ 93d达到 1 0 0 %(44 4 4) ;SARS CoVIgG第 1周检出阳性率为 4 .31 %(5 1 1 6 ) ,第 2周为 5 5 .88%(38 6 8) ,第 3周为 77.2 7%(1 7 2 2 ) ,2 2～ 93d达到 1 0 0 %(44 4 4) ;临床疑似病例N IgG检出阳性率为 3.1 9%(6 1 88) ,SARS CoVIgG检出阳性率为 2 .6 6 %(5 1 88) ;临床排除病例N IgG和SARS CoVIgG检出阳性率均为 6 .4 5 %(4 6 2 )。两种方法检测阳性率差异无统计学意义 (χ2 =0 .1 80 ,P <0 .0 0 1 ) ;两者之间符合率为 98.2 0 %(491 5 0 0 ) ,正常人群SARS CoVN IgG阳性率为 1 .88%(1 4 74 5 )。结论　SARS CoVN重组蛋白对于SARS诊断试剂的研究具有重要价值。     

囊尾蚴病免疫机理的研究现状

人们发现以链状带绦虫卵再次攻击感染家猪后,其体内的囊尾蚴逐渐消失;再次重复感染后,能加速囊尾蚴的变性与吸收。根据这一现象,Herbert 等首先提出免疫反应可能参与消除猪囊尾蚴。随后几年的研究结果显示,在囊尾蚴与宿主的关系中,免疫应答可能起关键作用。一、抗体在抗囊尾蚴感染中的作用Flisser 证明,在大约50%囊尾蚴病人的血清中含有高浓度的循环抗体。在多数脑、眼囊尾蚴病患者的脑脊液和房水中及其他类型患者的血清中,发现了IgG、IgM、IgA 和IgE 4种抗囊尾蚴抗体。患者血清中,占优势的抗体为IgG,约占全部病例的98%。以免疫电泳法检测特异性IgE 的阳性率为37%。初次注射囊尾蚴抗原一周后,实验动物的抗体水平开始升高,6～7周达高峰。虽然注射抗原后IgM 和IgG 水平都升高,但IgG 增高较明显。为了证实浆细胞     

SARS-CoV核衣壳蛋白抗原性鉴定和基因免疫研究

目的　了解SARS冠状病毒 (SARS CoV)核衣壳蛋白 (N蛋白 )的抗原性及其基因疫苗的免疫原性。方法　用大肠杆菌表达SARS CoV的N蛋白 ,用SARS患者恢复期血清对其进行抗原性鉴定。再构建N蛋白基因疫苗 ,肌肉注射接种小鼠 ,检测小鼠血清中的抗N蛋白IgG抗体、脾细胞增殖和迟发型超敏反应。结果　大肠杆菌重组表达的SARS CoVN蛋白具有强抗原性 ,其基因疫苗能在小鼠有效诱导N蛋白特异性抗体和CD4 、CD8 T淋巴细胞免疫应答。结论　SARS CoV的N蛋白可作为重要的SARS血清学诊断抗原 ,并对于SARS的特异性预防和治疗有潜在的应用价值。     

脆弱类杆菌NCTC9343脂多糖抗体的产生

本实验用间接血凝方法检测了兔抗血清中抗脆弱类杆菌NCTC 9343株脂多糖(LPS)抗体的变化情况。在初次免疫应答中,特异性抗体产生得很快,免疫局第2天即可测出,持续3～4周,高峰出现在第6～8天,最高效价为1:80;再次免疫应答,抗体高峰出现在再次免疫后的2～4天,效价为1:320,抗体效价维持6～8周。特异性抗体的类型主要是IgM。     

孕妇及其新生儿感染HCMV-IgG、IgM的检测分析

目的了解孕妇及其新生儿感染人巨细胞病毒（HCMV）的状况。方法采用ELISA法对200例孕妇及其新生儿进行HCMV特异性IgG和IgM抗体检测。结果孕妇血清中IgG和IgM抗体阳性率分别为96.5%和28.5%;新生儿脐血中IgG和IgM抗体阳性率分别为46.5%和11.5%;在23例异常产儿中其母亲HCMV特异性IgM抗体均为阳性。结论孕妇有HCMV近期感染对新生儿有一定影响。     

严重急性呼吸综合征患者IgG抗体的研究

目的　探讨血清特异性IgG抗体检测对严重急性呼吸综合征 (SARS)临床诊断的意义及影响因素。方法　用华大公司研制的SARS冠状病毒抗体 (IgG)酶联免疫试剂盒 ,对部分住院SARS患者的血清进行IgG抗体检测 ,统计分析不同年龄、性别和激素使用患者的SARS特异性抗体的产生情况。结果　在确诊的 12 1例患者中 ,IgG抗体阳性率为 71 1% ,年龄 <15岁、15～ 5 9岁和 6 0岁以上的患者 ,阳性检测率分别为 6 0 0 %、70 2 %和 85 7% ,差异无显著意义 (P =0 76 6 ) ;使用激素和不使用激素的患者 ,阳性率分别为 70 6 %和 72 4 % (P =0 84 ) ;病重和病情较轻患者的阳性检测率分别为78 1%和 6 7 4 % (P =0 4 93) ;男性和女性患者检测率均为 71 1%。结论　SARS特异性IgG抗体检测的敏感性有待进一步提高 ,IgG抗体的产生与性别、年龄、病情轻重和激素的使用无显著性关系。     

Recombinant truncated nucleocapsid protein as antigen in a novel immunoglobulin M capture enzyme-linked immunosorbent assay for diagnosis of severe acute respiratory syndrome coronavirus infection

We report the development of an immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) for severe acute respiratory syndrome coronavirus (SARS-CoV) by using recombinant truncated SARS-CoV nucleocapsid protein as the antigen. The newly developed MAC-ELISA had a specificity and sensitivity of 100% as evaluated by using sera from healthy volunteers and patients with laboratory-confirmed SARS. Using serial serum samples collected from SARS patients, the times to seroconversion were determined by IgM antibody detection after SARS-CoV infection. The median time to seroconversion detection was 8 days (range, 5 to 17 days) after disease onset, and the seroconversion rates after the onset of illness were 33% by the first week, 97% by the second week, and 100% by the third week. Compared with the results of our previous report on the detection of IgG, the median seroconversion time by IgM detection was 3 days earlier and the seroconversion rate by the second week after the illness for IgM was significantly higher than by IgG assay. Our results indicating that the IgM response appears earlier than IgG after SARS-CoV infection in consistent with those for other pathogens. Our newly developed MAC-ELISA system offers a new alternative for the confirmation of SARS-CoV infection.     

Chronological evolution of IgM, IgA, IgG and neutralisation antibodies after infection with SARS-associated coronavirus

Abstract Serum levels of IgG, IgM and IgA against severe acute respiratory distress syndrome (SARS)-associated coronavirus (SARS-CoV) were detected serially with the use of immunofluorescent antibody assays in 30 patients with SARS. Seroconversion for IgG (mean 10 days) occurred simultaneously, or 1 day earlier, than that for IgM and IgA (mean 11 days for both). IgG could be detected as early as 4 days after the onset of illness. The earliest time at which these three antibodies reached peak levels was similar (mean 15 days). A high IgG level (1:800) could persist for > 3 months. The kinetics of neutralisation antibodies obtained with 100x the tissue culture infective dose (TCID50) of the SARS-CoV TW1 strain in five patients with SARS nearly paralleled those for IgG. There were no significant differences in the kinetics of the IgG, IgM and IgA responses between patients with or without underlying medical disease, steroid or intravenous immunoglobulin therapy, or mechanical ventilation.     

Measles antibodies in nasal secretions and sera of children with measles.

Measles antibodies were determined, in the course of measles, in sera and nasal secretions of 54 and 27 children, respectively. The examinations were performed on the 3rd or 4th day (1st period) and between the 10th and 14th day (2nd period) after onset of rash in both sera and nasal secretions and after 25 to 60 days (3rd period) in sera only. Geometric mean titres of antibodies in sera determined by haemmagglutination inhibition (HI) and neutralization tests in the 1st period were 126.3 and 115.3 respectively. For the 2nd period the respective figures were 318.4 and 396.1 and for the 3rd period--388.0 and 445.6. Fractionation on Sephadex G-200 of sera from the 1st period revealed HI and neutralizaing antibodies associated mainly with the IgM serum globulin class. Measles HI and neutralizing antibodies were also found in nasal secretions of all 27 children, but their titres were much lower than in serum. The antibodies determined by indirect immunofluorescence in nasal secretions were associated with IgA in 26 and with IgG immunoglobulin in 15 of the 27 subjects. No IgM antibodies were found.     

Immunogenicity of SARS inactivated vaccine in BALB/c mice

Severe acute respiratory syndrome (SARS) is a serious infectious threat to public health. To create a novel trial vaccine and evaluate its potency, we attempted to generate a SARS inactivated vaccine using SARS coronavirus (SARS-CoV) strain F69 treated with formaldehyde and mixed with Al(OH)3. Three doses of the vaccine were used to challenge three groups of BALB/c mice. We found that the mice exhibited specific IgM on day 4 and IgG on day 8. The peak titers of IgG were at day 47 in low-dose group (1:19,200) and high-dose group (1:38,400) whereas in middle-dose group (1:19,200), the peak was at day 40. On day 63, the IgG levels reached a plateau. Neutralization assay demonstrated that the antisera could protect Vero-E6 cells from SARS-CoV's infection. Analysis of the antibody specificity revealed that the mouse antisera contained a mixture of antibodies specifically against the structure proteins of SARS-CoV. Furthermore, the mouse antisera conferred higher amount of antibodies against protein N, polypeptide S4 and S2 than those of proteins M and 3CL. These findings suggest that the inactivated SARS-CoV could preserve its antigenicity and the inactivated vaccine can stimulate mice to produce high levels of antibodies with neutralization activity. Results also suggest that polypeptides originating from protein N or S might be a potential target for the generation of a recombinant SARS vaccine.     

Persistent shedding of viable SARS-CoV in urine and stool of SARS patients during the convalescent phase

In order to further the present knowledge of the emerging severe acute respiratory syndrome-associated coronavirus (SARS-CoV), 486 different specimens from 54 patients with a clinical diagnosis of SARS were investigated for the presence of viral RNA, and 314 plasma specimens of 73 patients were examined for IgM and IgG antibodies specific against SARS-CoV using an indirect ELISA. Viral RNA was detectable in 28 of the 54 patients tested. Cumulative data showed that 67 of the 73 SARS patients demonstrated seroconversion by week 5 of illness. In contrast, only 1 of 278 healthy subjects enrolled in the study was found to be positive for the IgG antibody. Coexistence of viral RNA in plasma and specific antibodies was simultaneously observed over three consecutive weeks in two critical cases. In three convalescent patients in particular, cultivable SARS-CoV was detected in stool or urine specimens for longer than 4 weeks (29–36 days). These findings suggest that SARS-CoV may remain viable in the excretions of convalescent patients.     

人冠状病毒229E、OC43与SARS-CoV血清学相关性的研究

目的 检测正常人和SARS患者血清中3种人冠状病毒（229E、OCA3和SARS-CoV）特异性抗体．分析3种冠状病毒血清学相关性。方法 采用免疫印迹、免疫荧光和ELISA方法检测100例健康献血员、34例SARS患者恢复期以及11例SARS患者双份血清中229E、OCA3和SARS-CoV3种冠状病毒核衣壳（N）蛋白抗体。结果 用免疫荧光方法检测100例健康献血员血清中229E、OCA3和SARS-CoV IgG阳性率分别为98％、100％和1％，34例SARS患者恢复期血清中3种冠状病毒IgG的阳性率均为100％；免疫印迹检测100例健康献血员血清中229E、OCA3和SARS-CoVN蛋白IgG阳性率分别9r7％、99％和2％，34例SARS患者恢复期血清中229E、OCA3和SARS-CoVN蛋白IgG阳性率分别97％、100％和100％；11例SARS患者的急性期和恢复期双份血清中，免疫荧光检测有5例出现229E IgG滴度4倍或以上升高，10例出现OC43 IgG滴度4倍或以上升高，ELISA检测2例出现229EN蛋白IgG滴度4倍以上升高，没有一例出现OCA3N蛋白抗体滴度升高。结论 正常人群中普遍存在229E和OCA3两种人冠状病毒抗体，SARS-CoV感染者存在对人冠状病毒229E和OCA3血清学交叉反应，提示核衣壳蛋白不是引起血清学交叉反应的主要抗原，结果对研究SARS溯源有重要意义。     

Cross-reaction of SARS-CoV antigen with autoantibodies in autoimmune diseases

To investigate the significance of the SARS-associated coronavirus (SARS-CoV) antibody,detected by ELISAand indirect immunofluorescence assays (IFA) for the SARS-CoV Vero E6 cell lysates,in non-SARS subjects,114 serum samples from healthy controls and 104 serum specimens from autoimmune disease patients werecollected.The results of ELISA showed that among 114 sera from healthy controls,4 (3.5%) were positive ofSARS-CoV-IgG antibody and 114 (100%) were all negative of SARS-CoV-IgM antibody;the specificity ofSARS-CoV-IgG antibody for SARS patients was 96.5%,but the specificity of both SARS-CoV-IgG and -IgMantibodies for SARS patients was 100%.In 58 cases with SLE,positive rates of SARS-CoV-IgG and -IgMantibodies were 32.8% (19/58) and 8.6% (5/58),respectively,in which 11 cases (19%) were positive of bothSARS-CoV-IgG and -IgM antibodies;in 10 cases with SS,positive rate of both SARS-CoV-IgG and -IgMantibodies was 10% (1/10);in 16 cases with MCTD,positive rate of SARS-CoV-IgG was 37.5% (6/16),positiverate of both SARS-CoV-IgG and -IgM antibodies was 6.3% (1/16);in 20 cases with RA,one case was positive(5%) of SARS-CoV-IgG.However,of all samples with positive SARS-CoV-IgG and -IgM antibodies forautoimmune diseases and healthy controls,SARS-CoV RNA and antibodies were all negative by RT-PCR andIFA.All sera for negative or positive ELISA results were also negative or positive results using ELISA withVero E6 cells lysates.These studies showed that SARS-CoV Vero E6 cell lysates for the ELISA to detectSARS-CoV antibodies could lead to the false-positive reactions or cross-reactions of SARS-CoV antibodies innon-SARS diseases and healthy controls,and the false-positive reactions or cross-reactions were related to VeroE6 cell lysates and autoantibodies in non-SARS population.Cellular & Molecular Immunology.2004;1(4):304-307.     

Detection of the nucleocapsid protein of severe acute respiratory syndrome coronavirus in serum: comparison with results of other viral markers

A capture enzyme-enhanced chemiluminescence immunoassay (ECLIA) based on three specific monoclonal antibodies to detect the nucleocapsid (N) protein of severe acute respiratory syndrome (SARS) associated coronavirus (SARS-CoV) in the serial serum samples from SARS patients was developed. The anti-SARS-CoV IgG and the viral RNA were also detected in the sera by ELISA and RT-PCR, respectively. During the first 10 days after onset, anti-SARS-CoV IgG, SARS-CoV RNA and the N protein were detected in 21.4, 42.9, and 90% of the patients’ sera, respectively. The detection rate of the N protein during days 11–15 of the disease was still significantly higher than those of anti-SARS-CoV IgG and SARS-CoV RNA. The data demonstrated that detection of the N protein with the capture ECLIA appears to be more useful than detection of other viral makers for rapid diagnosis of SARS in patients.