天津某医院严重急性呼吸综合征暴发期间血清流行病学调查

目的 研究本院严重SARS暴发流行后，疫源地不同人群血清SARS冠状病毒（SARS-CoV）抗体水平及其产生和变化规律；探讨人群SARS-CoV隐性感染和IgG抗体的保护作用。方法 采用血清流行病学法，联合运用ELISA和免疫荧光试验（IFA），调查疫情暴发流行期间及流行后，疫源地内非SARS人群血清抗体水平变化；定性研究SARS患者病后6周内IgM、IgG抗体产生和变化规律；动态观察SARS患者康复期82周内IgG抗体水平变化规律。结果 各100例疫源地一般人群和非疫源地对照人群血清SARS-CoV IgG抗体ELISA抽样检测均为阴性；487例SARS高危人群血清SARS-CoV IgG抗体ELISA检测阳性率为0．41％，经IFA复核后均为阴性；疫源地内非SARS人群IgG抗体A值水平在疫情流行后高于流行期间，差异有统计学意义（P〈0．05）；SARS患者病后1～6周血清抗体IgM和IgG的阳性检出率分别为7．7％、40．0％、57．8％、88．2％、76．6％、57．1％和0、23．1％、48．4％、65．4％、77．8％、100．0％；SARS患者康复期血清IgG抗体A值逐渐增高，病后第22周达到高峰，此后缓慢下降，第82周时仍维持较高水平，下降趋势减缓。结论 SARS可能不存在隐性感染者；绝大多数SARS患者在急性期或恢复早期的血清中存在IgM抗体，其出现较早，消失也较快，抗体IgG出现稍晚，但在血清中存在时间较长，达到高峰后消退缓慢，提示该抗体可能具有保护作用。     

传染性非典型肺炎病例SARS冠状病毒特异性抗体检测及分析

目的　对福建省传染性非典型肺炎病例血清标本中SARS冠状病毒 (SARS -CoV)特异性抗体进行检测。方法　采用间接酶联免疫吸附试验 (ELISA)测定SARS临床诊断、疑似、医学观察病例以及其他非SARS患者和健康人群血清SARS -CoV特异性IgM、IgG抗体。 结果　 (1) 3例SARS临床诊断病例血清SARS -CoV特异性IgG抗体均为阳性 ,其中 2例IgM抗体阳性 ,2例恢复期血清比急性期血清IgG抗体呈≥ 4倍增长 ;(2 ) 1例疑似病例IgM和IgG抗体均为阳性 ,且恢复期血清比急性期血清IgG抗体呈 >4倍增长 ;(3) 11例医学观察病例IgM和IgG抗体均为阴性 ;(4 ) 5 0例非SARS患者和健康人群IgG抗体全部阴性。 结论　间接ELISA是传染性非典型肺炎实验室特异性检测方法 ,可对SARS临床诊断进行进一步的核实。     

严重急性呼吸综合征相关性冠状病毒抗体检测及分析

目的 了解发病期严重急性呼吸综合征(SARS)患者体内Urbani SARS相关性冠状病毒抗体产生的规律，探讨这些患者中肺炎衣原体、肺炎支原体、腺病毒、呼吸道合胞病毒感染情况。方法 采用免疫荧光技术检测43例SARS患者双份血清和10例非SARS患者血清中Urbani SARS冠状病毒抗体IgM／IgG，并采用酶联免疫法检测上述血清中肺炎衣原体、肺炎支原体、腺病毒、呼吸道合胞病毒IgM／IgG。结果 43例患者中新近感染Urbani SARS冠状病毒40例(93％)，3例未检出其感染；10例非SARS患者血清中Urbani SARS冠状病毒抗体IgM／IgG均为阴性。肺炎衣原体、肺炎支原体、腺病毒、呼吸道合胞病毒新近感染率分别为25．6％、163％、7％和4．7％，其既往感染率分别为39．5％、34．9％、27．9％和0。SARS病毒IgM抗体在起病开始便能检测到，其阳性率在起病8～14d和15～33d时分别为69．6％和63％，二者比较无统计学差异，但均显著高于起病1～7d时的阳性率(16．7％)；SARS病毒IgG抗体在起病6d时便能检测到，其阳性率在起病1～7d、8～14d和15～33d时分别为19．4％、65．2％和92．6％，三者之间比较均差异有显著性。结论 SARS患者发病初期即可出现Urbani SARS冠状病毒IgM和IgG抗体，发病2周至1个月时检测阳性率明显增加。部分SARS患者中存在肺炎衣原体、肺炎支原体、腺病毒、呼吸道合胞病毒新近感染和／或既往感染。免疫荧光技术榆测Urbani SARS冠状病毒抗体可用于SARS的早期诊断。     

SARS患者及其医护人员血中特异IgG抗体变化规律的初步研究

用酶联免疫吸附测定(ELISA)检测57例正常人、127例在SARS病房工作1个月的医护人员和73例不同病程SARS患者血浆抗SARS冠状病毒IgG抗体的水平。结果:正常人和医护人员血浆中尚未检测到抗SARS冠状病毒IgG抗体。SARS患者发病第0-7、8-10、11-14、15-20天后血浆中抗SARS冠状病毒IgG抗体的阳性率分别为0、33％、52％和86％,总阳性率为61％。提示ELISA检测血浆中抗SARS冠状病毒IgG抗体的特异     

杆状病毒表达SARS冠状病毒核蛋白抗原性的研究

目的 对杆状病毒表达SARS冠状病毒NP抗原性进行分析与比较。方法 利用SDS—PAGE、Western—Blot和ELISA证明重组核蛋白(rSN)在昆虫细胞获得高效表达．具有特异免疫反应原性。结果 表达rSN的昆虫细胞裂解、稀释后直接包被ELISA板．来检测SARS-CoV康复病人血清特异抗体，敏感性稍高于Veto细胞培养的全病毒(OD分别为2．8和0．6)。与健康人血清不发生特异反应(OD值为0．01)；表达rSN昆虫细胞用于间接免疫荧光，快速检测血清特异抗体反应。同样表现了良好的敏感性和特异性。结论昆虫细胞表达rSN有望替代SARS-CoV全病毒，作为安全、敏感和特异的诊断抗原。应用于ELISA和IFA血清学检测。     

抗核抗体不同检测方法的对比分析

目的评价以HEp-2细胞为底物的间接免疫荧光法(IFA)和包被多种纯化核抗原的酶联免疫吸附试验(ELISA)检测抗核抗体(ANA)的特点及其在系统性红斑狼疮(SLE)诊断中的应用价值。方法同时采用IFA法和ELISA法检测血清ANA共409例,其中SLE 226例和健康对照183名,将两种方法检测结果的敏感度、特异度、准确度、阳性预测值、阴性预测值、阳性似然比、阴性似然比、结果符合率、判断一致性κ值及秩相关系数进行比较分析；通过ROC曲线比较两种方法的准确度；结果不一致标本进行可提取性核抗原抗体谱(ENA)检测分析；同时将IFA法检测不同荧光模型样本分别与ELISA结果进行相关分析。结果SLE患者组IFA法和ELISA法检测ANA的敏感度分别为91．15%和92．04%,特异度分别为96．17%和92．90%,准确度分别为93．40%和92．42%,阳性预测值分别为96．71%和94．12%％,阴性预测值分别为89．80%和90．43%．两种方法比较差异均无统计学意义(P＞0．05),ROC曲线对两种方法准确度分析差异无统计学意义(P=0．409)；两方法检测结果符合率为91．20%,秩相关系数R=0．823,判断一致性κ=0．825：在36例两种方法检测结果不一致标本中,14例IFA( )ELISA(一),其中1例IFA法滴度达1：1000的高尔基体型．其余为临界值左右的核仁型和斑点型,而在22例IFA(一)ELISA( )患者中,其中有11例ELISA法吸光度(A)2．67～30．5；将此36例血清标本进行ENA抗体谱检测,14例IFA( )ELISA(一)患者ENA阳性率为14．29%,而22例IFA(一)ELISA( )患者ENA阳性率高达68．18%,两者比较差异有统计学意义(P＜0．01)。IFA法检测的不同荧光模型的阳性标本中,斑点型滴度与ELISA法A值相关性较差R=0．083,均质型相关性最好。R=0．595。结论IFA作为ANA检测的推荐方法具有直观、可分型等特点,且能提供与自身免疫性疾病相关的核型信息,但需要荧光显微镜及较丰富判断经验的技术人员；而ELISA法操作简单。并可通过测定A值来评价患者体内抗体的浓度,不需再作滴度检测；IFA法和ELISA法检测ANA均有较高的敏感度、特异度、准确性,结果符合率、一致性判断系数κ和秩相关系数均较高,而后者的漏诊率更低．更适合进行ANA筛查；若对ELISA板再增加包被高尔基体、核仁等稀有荧光模型的抗原,则可使ELISA法检测结果更准确、更全面,对申请ANA检测的标本,可先采用ELISA法筛选出阳性标本后再进行IFA法检测,这一工作流程可加快报告速度,节约检验成本,提高工作效率。     

203名吸毒人员血液和尿液HIV抗体检测结果对比分析

目的 评价尿液艾滋病病毒(HIV-1)抗体检测方法。方法 用荷兰阿克苏检测血液和美国卡里普特检测尿液HIV-1抗体的酶联免疫吸附试验(ELISA)试剂，分别同时检测吸毒人员的血液和尿液中HIV-1抗体，并与血液检测结果比较，计算尿液检测方法的敏感度、特异度和一致性。结果 检测203名吸毒人员，血液HIV-1抗体阳性127人，尿液HIV-1抗体阳性125人，尿液检测阳性敏感度为98．4％，特异度为100％，一致性为99．0％。结论 尿液HIV抗体检测结果与血液检测结果有较好的一致性。     

用Sephadex G200层析血清对4种弓形虫病免疫诊断方法的评价

弓形虫感染小鼠血清经Sephadex G 200柱层析后,各层析峰(紫外线OD_(280)吸收峰)用染色试验(DT)、乳胶凝集试验(LAT)、间接荧光抗体试验(IFA)和酶联免疫吸附试验(ELISA)分别检测总抗体、IgM和IgG抗体。结果显示,两类抗体分别存在于第Ⅰ峰和第Ⅱ峰,第Ⅲ峰无抗体活性。ELISA和IFA用于弓形虫病早期诊断(检测IgM)具有高度特异性和敏感性;LAT操作简便、经济快速、但敏感性稍差。IFA可取代DT用于常规个例诊断。     

酶联免疫吸附试验检测人血清弓形体IgG,IgM抗体的研究

对ELISA检测人血清弓形体IgG、IgM抗体进行了研究。450份孕妇血清中,ELISA阳性率显著高于IHA;抗体滴度分折,ELISA一般高于IHA2～10倍。25份ELISA IgG抗体阳性血清,IFA检出19份;3份IgM抗体阳性和2份IgG、IgM抗体均阳性血清,IFA分别检出2份。2份阴性和4份含不同抗体滴度的阳性血清于第一次测定后,第7天和第21天测定的阴、阳性结果一致,OD值变异系数为2.43～16.52％。39份阳性血清抗体滴度与OD值呈直线比例关系(r=0.991,P<0.0005)。结果表明,ELISA用于弓形体感染的血清学诊断具有较好的实用性。     

血清新型冠状病毒特异性抗体水平的检测及其动态变化规律

重症急性呼吸综合征(SARS)的病原体相继被不同实验室发现并确证为一种新型冠状病毒，又称为SARS病毒，已确定了病毒基因的一级结构和一些基因组特征，为利用分子免疫学手段研究和建立SARS的血清学诊断方法奠定了基础。我国军事医学科学院等单位已成功建立了检测SARS病毒抗体的ELISA方法，并开发出相应的检测试剂盒。本研究采用ELISA方法检测SARS患者和医务人员血清SARS病毒特异IgG抗体，观察SARS病毒抗体在患者和高危易感人群的产生规律和水平，为SARS的诊断及其流行传播特点的分析提供实验依据。     

Serodiagnosis of Rocky Mountain spotted fever: comparison of IgM and IgG enzyme-linked immunosorbent assays and indirect fluorescent antibody test

To characterize IgM and IgG antibody responses in Rocky Mountain spotted fever (RMSF), a microtiter enzyme-linked immunosorbent assay (ELISA) using density gradient-purified Rickettsia rickettsii as antigen was developed. Sera of vaccinated individuals and patients with RMSF were tested by ELISA and by indirect fluorescent antibody (IFA) tests. Diagnostic agreement between ELISA and the IFA test was 76% and 52% for IgG and IgM antibody, respectively. Diagnostic agreement between the ELISA for IgG antibody and the IFA test for total immunoglobulins was 84%. The ELISAs for IgM and IgG antibody were as specific (100%) and as sensitive (100%) as the IFA test (83%-100%) in detecting antibody increases in paired sera from persons with RMSF and were superior to the IFA test in detecting seroconversions in vaccinees. The ELISA also detected antibodies in a single convalescent-phase serum with sensitivity and reliability. The ELISA for IgG antibody is appropriate for seroepidemiology and serodiagnosis since it permits measurement of antibody at a single dilution of serum up to a year after illness.     

检测华支睾吸虫特异性IgG4生物素-亲和素复合酶联免疫吸附法的建立及检测效能分析

目的探讨亲和素-生物素复合酶联免疫吸附法(ABC-ELISA)检测特异性Ig G4抗体用于诊断华支睾吸虫病的效果。方法建立检测华支睾吸虫特异性抗体Ig G4的ABC-ELISA法(Ig G4-ABC-ELISA),以此方法检测华支睾吸虫病、日本血吸虫病、卫氏并殖吸虫病、弓形虫病、棘球蚴病、囊尾蚴病和曼氏裂头蚴病患者血清样本。以Ig G4-ELISA和Ig GELISA法为对照,比较3种方法用于诊断华支睾吸虫病的敏感性、特异性、阳性预测值、阴性预测值及诊断效率。结果成功建立了用于检测华支睾吸虫特异性抗体的Ig G4-ABC-ELISA法,用此方法检测华支睾吸虫病患者血清特异性抗体Ig G4的敏感性为90.0%,特异性为98.2%,阳性预测值为93.8%,阴性预测值为97.0%,诊断效率为96.3%;Ig G4-ELISA法检测华支睾吸虫病患者血清特异性抗体敏感性为86.0%,特异性为98.2%,阳性预测值为93.5%,阴性预测值为95.9%,诊断效率为95.4%;Ig G-ELISA法检测华支睾吸虫病患者血清特异性抗体Ig G的敏感性为94.0%,特异性为88.1%,阳性预测值为70.1%,阴性预测值为98.0%,诊断效率为89.4%。Ig G4-ABC-ELISA法检测华支睾吸虫病患者血清特异性抗体的敏感性高于Ig G4-ELISA法(P0.05),特异性高于Ig G-ELISA法(P0.05)。结论 Ig G4-ABC-ELISA法检测华支睾吸虫特异性抗体IgG4具有高度敏感性与特异性,在华支睾吸虫病诊断中具有较好应用价值。     

严重急性呼吸综合征患者冠状病毒总抗体及N和S蛋白抗体的随访研究

目的了解SARS冠状病毒（SARS—CoV）感染者血清中特异性IgM、IgG抗体和两个结构蛋白抗体的持续时间及相互关系。方法对146例SARS临床确诊且血清抗-SARS—CoV阳性病例,随访采集发病当13至发病后660d期间的血液标本共362份。以ELISA法分别检测抗-SARS—CoVIgM和IgG抗体、SARS—CoVN蛋白和S蛋白IgG抗体。结果抗-SARS—CoV IgM阳性率在发病20d内为46．5％（20／43）,21—40d阳性率最高（80．6％,25／31）,尔后逐渐下降,至发病后500d左右仅为8．2％（6／73）。IgG总抗体在发病20d内阳性率（34．9％,15／43）低于IgM抗体,在发病21～40d迅速达到100％,至发病后600—660d,其阳性率仍高达98．6％（70／71）。N—IgG抗体在发病40d后阳性率（92．5％,37／40）高于S-IgG（67．5％,27／40）,在61～90d、450—510d和600—660d3个时间点检测阳性率均高于S-IgG抗体;但两种结构蛋白抗体阳性率随时间延长均逐渐下降,两者于不同时间点的阳性率均低于抗-SARS—CoV总IgG。结论临床与病原学确诊的SARS患者,SARS—CoV特异性抗体阳性率在21—40d达高峰,IgG抗体阳性率100％。IgM抗体91．8％在感染500d以内消失;感染后两年IgG总抗体阳性率仍高达98．6％,推测可持续阳性3—5年。N—IgG和S-IgG抗体持续时间可能较短。     

Longitudinal profile of antibodies against SARS-coronavirus in SARS patients and their clinical significance

BACKGROUND: Severe acute respiratory syndrome (SARS) is a newly discovered disease caused by a novel coronavirus. The present study studied the longitudinal profile of antibodies against SARS-coronavirus (SARS-CoV) in SARS patients and evaluated the clinical significance of these antibodies. METHODS: Two methods, ELISA and indirect immunofluorescent assay, were used for the detection of the anti-SARS-CoV IgG and IgM in 335 serial sera from 98 SARS patients. In 18 patients, serum antibody profiles were investigated and antibody neutralization tests were performed from 7 to 720 days after the onset of symptoms. RESULTS: The ratios of positive IgG/IgM by ELISA were 0/0, 45.4/39.4, 88.6/71.4, 96/88, 100/48.6, 100/30.9, 100/17.1, 100/0 per cent, respectively, on 1-7, 8-14, 15-21, 22-28, 29-60, 61-90, 91-180 and 181-720 days after the onset of symptoms. Antibodies were not detected within the first 7 days of illness, but IgG titre increased dramatically on day 15, reaching a peak on day 60, and remained high until day 180 from when it declined gradually until day 720. IgM was detected on day 15 and rapidly reached a peak, then declined gradually until it was undetectable on day 180. Neutralizing viral antibodies were demonstrated in the convalescence sera from SARS patients. CONCLUSION: The persistence of detectable IgG antibodies and neutralizing viral antibodies for up to 720 days suggest that SARS patients may be protected from recurrent SARS-CoV infection for up to 2 years.     

SARS-CoV抗体实验诊断的特异性研究

为探讨严重急性呼吸综合征(SARS)病毒(SARS-CoV)抗体在SARS病原学诊断中的特异性，应用酶联免疫吸附试验(ELISA)和荧光定量RT-PCR技术检测了80例非SARS患者SARS-CoV抗体的阳性率。结果在23例健康人中，SARS-CoV-IgG抗体的阳性率为8．7％(2／23)，20例肿瘤患者中，阳性率为20％(4／20)；18例自身免疫性疾病患者中，SARS-CoV-IgM抗体和SARS-CoV-IgG抗体阳性率分别为11．1％(2／18)和22．2％(4／18)；19例系统性红斑狼疮(SLE)患者中，SARS-CoV-IgM抗体和SARS-CoV-IgG抗体阳性率分别为21．1％(4／19)和47．4％(9／19)，SARS-CoV-IgG抗体和SARS-CoV-IgM抗体同时阳性率为15．8％(3／19)。证实SARS-CoV-IgM抗体诊断SARS的特异性为92，5％；SARS-CoV-IgG抗体诊断SARS的特异性为76．25％；两种抗体同时阳性诊断SARS的特异性为96．25％。经RT-PCR证实上述抗体单项阳性均为假阳性。认为两种抗体同时测定可提高诊断的特异性，出现假阳性的原因可能与包被的抗原中存在细胞膜、胞浆、核和抗细胞核成分的自身抗体有关。     

Clinic–Epidemiologic Study of Human Infection by Granada Virus,a New Phlebovirus within the Sandfly Fever Naples Serocomplex

Granada virus (GRV), a new phlebovirus within the Naples serocomplex, has been recently described in phlebotomine sandflies from Spain. The presence of anti-GRV immunoglobulin G (IgG) antibodies was investigated by indirect fluorescence assay (IFA) and neutralization test (NT) in 920 serum samples from the Granada population. By IFA, an overall GRV seroprevalence of 15.8% (N = 145) was observed, significantly increasing up to 65 years. NT was positive in 18% of anti-GRV IFA-positive samples. IgG antibodies against Toscana virus (TOSV), a hyperendemic phlebovirus within Granada province, were detected in 40% of anti-GRV–positive cases. Anti-GRV IgM antibodies were detected in 36 (6.6%) of 547 acute-phase serum samples from individuals with febrile illness, exanthema, and/or acute respiratory infection. All positives were anti-TOSV IgM-negative. GRV may infect humans, with most cases being asymptomatic. The codetection of anti-GRV and anti-TOSV IgG antibodies could be attributable to cross-reactivity or exposure to the same transmission vector.     

Incidence of primary dengue virus infections in Southern Vietnamese children and reactivity against other flaviviruses

Objective To study the incidence of asymptomatic primary dengue infections among children and reactivity against other flaviviruses. Methods A total of 216 children, who had no dengue‐specific IgG antibodies during a serosurvey in 2003 were re‐examined 23 months later to determine if seroconversion had occurred. Dengue‐specific IgG was demonstrated with enzyme‐linked immunosorbent assay (ELISA) and reactivity patterns against other flaviviruses were assessed by using immunofluorescence assay (IFA). Results Sixty‐six children had seroconverted for dengue virus‐specific IgG; the true annual incidence of primary dengue was thus 17.3% (95% CI: 13.8–21.4). Japanese Encephalitis virus (JEV)‐specific IgG antibodies were detected by IFA among three (4.6%) samples that showed seroconversion in the dengue ELISA, because of cross‐reactivity. Conclusion Our findings highlight the high incidence of dengue among Vietnamese children; JEV infections are rare. The true annual incidence of dengue can be estimated with a single cross‐sectional seroprevalence survey.     

Evaluation of the clinical relevance of anti-annexin-A5 antibodies in Chinese patients with antiphospholipid syndrome

A hallmark feature of antiphospholipid syndrome (APS) is the presence of antiphospholipid antibodies (aPLs). Few studies have addressed the clinical relevance of anti-annexin A5 antibodies (aANXA5) in Chinese patients with APS. In this study, we evaluated the clinical performance of aANXA5 in the diagnosis of APS. Sera from 313 subjects were tested, including 170 samples from patients with APS, 104 samples from patients with non-APS diseases as disease controls (DC), and 39 healthy controls (HC). Serum IgG and IgM aANXA5 were determined by ELISA. Overall, the levels of both IgG and IgM aANXA5 were significantly increased in patients with primary APS (PAPS) and APS associated to other diseases (APSAOD) compared with DC and HC. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for IgG and IgM aANXA5 in the diagnosis of APS were 33.5 and 15.3, 99.0 and 99.0, 98.3 and 96.3, and 47.7 and 41.7%, respectively. Significant associations between IgG aANXA5 and arterial thrombotic events (OR, 2.60; 95% CI, 1.44–4.71) and between IgG aANXA5 and venous thrombotic events (OR, 2.80; 95% CI, 1.55–5.06) were identified. No correlations were identified between IgG or IgM aANXA5 and obstetric complications. Our data suggest that aANXA5 could serve as a diagnosis biomarker for patients with APS. More importantly, our data highlighted a potential role of IgG aANXA5 in identifying APS patients with high risk of thrombosis.