ELISA IFAT和PCR/生物素探针检测中枢神经系统疾病中弓形虫感染

目的　比较 ELISA、IFAT和 PCR/生物素探针检测弓形虫感染的效果 ,了解中枢神经系统疾病中弓形虫感染的情况。方法　应用 EL ISA、IFAT和 PCR/生物素探针 (PCR-BP)对 164例脑囊虫病、10 8例疑似脑囊虫病、88例原发性癫痫患者和 115例健康人群进行了血清弓形虫抗体及血液中弓形虫特异序列 DNA检测和比较。结果　 ELISA和 IFAT检测各组人群抗弓形虫抗体阳性率分别为 15 .85 %、14 .81%、13 .64 %和 6.0 9% ,14 .0 2 %、14 .81%、14 .77%和 6.0 9%。PCR-BP检测各组弓形虫特异性 DNA阳性率分别为 2 .44 % (4 /164 )、3 .70 % (4 /10 8)、3 .41% (3 /88)、0 (0 /115 )。结论　 PCR/生物素探针适合诊断现症弓形虫感染 ,而 EL ISA和 IFAT可作为弓形虫感染的筛选方法。脑囊虫病、疑似脑囊虫病和原发性癫痫患者中存在弓形虫感染     

脑囊虫病免疫诊断方法的比较

为了进一步比较皮内试验(ID)、IHA和ELISA在诊断脑囊虫病方面的应用价值,对136例经头颅CT诊断为脑囊虫病的患者进行了血清和脑脊液(CSF)免疫学诊断方法的比较。 检测对象 1 实验组 脑囊虫病患者136例中,皮下结节经活检证实为囊尾蚴者95例;单纯脑囊虫病者41例。     

用ELISA对吡喹酮治疗脑囊虫病的随访研究

作者采用ELISA对吡喹酮治疗后的脑囊虫病人进行血清学随访,以反映病灶的演变,评价其在监测吡喹酮治疗上的意义。 1984年1月至1986年6月,以经脑CT检查以CF为抗原,用ELISA检测血清和脑脊髓液(CSF)中的囊虫病IgG特异性抗体而确诊的69例脑囊虫病患者为观察对象,其中63例治疗前血清和/或CSF抗体阳性,其余6例两者均为阴性。采用吡喹酮3×25mg/kg 4、7和14天疗法,其中28例合并     

金标免疫渗滤法诊断疑似并殖吸虫病及考核疗效的观察

目的 探讨金标免疫渗滤法(DIGFA)检测并殖吸虫抗体的临床诊断价值。 方法 2003-2006年由浙江省有关医院和疾病预防控制中心转诊的疑似并殖吸虫病患者72例，用并殖吸虫抗体金标快速检测法(Pw-DIGFA）检测患者血清抗体阳性者，综合其饮食史及相关临床表现，拟诊为并殖吸虫病，给予吡喹酮治疗，并于治疗后3及6个月随访、复查，分别用Pw-DIGFA和ELISA检测患者治疗前、后配对血清抗体水平。 结果 Pw-DIGFA和ELISA 法检测结果一致，72例中抗体阳性28例，阴性44例。在28例阳性者均有生食或半生食淡水蟹或蝲蛄史，以及饮用生溪水史；21例有低热、咳嗽及胸肺部影像学改变等并殖吸虫病相关临床征状和体征，7例仅外周血嗜酸粒细胞异常增高（15％～70％）。用ELISA 检测阳性者、阴性者、健康人血清抗体水平(吸光度A₄₅₀值）分别为1.7361、0.2973、0.2657；经t检验，阳性者与阴性者间差异具有统计学意义（t =12.047, P< 0.01），阴性者与健康人之间差异无统计学意义（t=1.919, P>0.05）。治疗后3个月复查，ELISA检测6例中有5例血清效价下降2～5个滴度，Pw?鄄DIGFA检测抗体反应斑点色泽变淡；ELISA检测1例治疗后复发患者抗体效价升高1个滴度，Pw-DIGFA检测抗体反应斑点色泽变化不明显。治疗后6个月复查7例临床症状缓解或消失者, ELISA检测抗体效价下降3～6个滴度, Pw-DIGFA检测抗体反应斑点色泽较治疗前明显减弱。 结论 Pw-DIGFA检测疑似并殖吸虫病患者血清抗体有较好的临床辅助诊断价值, 治疗前、后配对血清平行检测有较好疗效考核价值。Pw-DIGFA法操作简易快速，适用于并殖吸虫病较少见地区的诊断和疗效评估。     

Dot-IGSS检测脑脊液中循环抗原诊断脑囊虫病的研究

应用斑点免疫金银染色法(Dot-IGSS)和Dot-ELISA检测96份确诊为脑囊虫病患者脑脊液(CSF)中的循环抗原(CAg),阳性率分别为94.79%和92.71%.检测51例非囊虫病者和62例其他脑部疾病患者的CSF,两种方法均为阴性.诊断指数分别为194.79%和192.71%,检测CAg的最小量Dot-IGSS为244ng/ml,Dot-ELISA为488ng/ml.试验表明两法诊断脑囊虫病均具有敏感性高、特异性强、重复性好和方法简便易行,有希望用于脑囊虫病的临床诊断.     

乳腺猪囊尾蚴病2例

例 1,女 ,36岁 ,工人 ,吉林市人。左乳腺组织内有 1cm×1.5 cm的肿块 ,触稍痛。市内医院以“乳腺纤维瘤”于 1999年12月手术摘除。病理诊断为乳腺猪囊尾蚴。翌年 5月因头痛、癫痫反复发作来我院。头部 CT诊断 :脑囊虫病。囊虫免疫检查 :CAg、IHA及 EL ISA均 ( )。综合诊断 :脑合并乳腺囊虫病。经住院抗囊虫治疗 ,临床治愈出院。例 2 ,女 ,2 7岁 ,农民 ,松原市人。主诉 5年前有绦虫病史。1999年夏 ,劳动时左乳房疼痛 ,触到 1cm× 1cm的结节 ,逐渐明显。经医院手术摘除 ,病理诊断为乳腺囊尾蚴病。囊虫免疫检查 :CAg、IHA及 EL ISA均…     

检测血清、脑脊液中循环抗原用于脑囊虫病诊断及疗效评估的研究

以猪囊尾蚴囊液抗原免疫家兔制备抗体,用常规ELISA法和斑点ELISA法(Dot-ELISA分别检测脑囊虫病人血清和脑脊液(CSF)中循环抗原(CAg),结果两法用于检测的敏感性与特异性相近。以Dot-ELISA法检测,脑囊虫病人血清和(CSF中CAg检出率分别为72.2％和77.7％,特异性为93.7％。吡喹酮治疗后,CSF中循环抗原消失较血清中明显缓慢,2疗程治疗后CSF中CAg检出率仍达74％,而血清中CAg检出率为40.7％,说明常规吡喹酮治疗后脑囊虫较外周组织中囊虫死亡缓慢,因此治疗后检测CSF中CAg更能判定疗效。     

间接ELISA检测弓形虫感染鼠NTPase-Ⅱ抗体的研究

目的建立检测小鼠血清中刚地弓形虫NTPase-Ⅱ特异性IgGI、gM抗体的间接ELISA法,探讨用于弓形虫感染早期检测的应用价值。方法以刚地弓形虫NTPase-Ⅱ融合蛋白为包被抗原建立间接ELISA法,动态检测刚地弓形虫实验感染鼠血清NTPase-Ⅱ特异性IgGI、gM抗体变化。试验设脑囊虫病、血吸虫病及肺吸虫病病人血清对照。结果小鼠血清中的抗NTPase-Ⅱ抗体IgM于弓形虫感染后第3 d开始出现阳性反应,IgG抗体于感染后第7 d开始出现阳性反应,脑囊虫病、血吸虫病及肺吸虫病病人血清特异性IgG阳性率分别为0、1.92%和0。结论间接ELISA检测弓形虫感染鼠血清NTPase-Ⅱ抗体的敏感性和特异性较高,有望应用于人感染弓形虫的早期检测。     

囊虫病的免疫诊断现状与展望

囊虫病是一种呈世界分布的人兽共患寄生虫病 ,对人类健康危害极大 ,尤其是脑囊虫病 ,如不及时治疗或治疗不当往往使人致残甚至危及生命。囊虫病的免疫学诊断可以弥补病原学、流行病学、影像学诊断的不足 ,在诊断和鉴别诊断中具有重要地位。近年来囊虫病的免疫学诊断发展很快 ,现将主要诊断方法和目前囊虫病诊断的工作重点及存在的主要问题综述如下。1　主要检测方法1.1　EL ISA及其改良的方法1.1.1　 Mc Ab- EL ISA　 EL ISA简单、灵敏 ,可批量测试 ,结果判定客观 ,是至今应用最广泛的一种免疫学诊断方法。单克隆抗体 (Mc Ab)具有特…     

脑瘤误诊为脑囊虫病9例分析

脑瘤和脑囊虫病均为占位性病变 ,临床表现有诸多共同症状 ,有的类型 CT、MRI检查和病理阶段与脑囊虫病有酷似的影像显示 ;脑瘤的血清和脑脊液囊虫病免疫学 IHA、EL ISA和CAg检查有假阳性出现 ,脑肿瘤性疾病中胶质瘤和转移瘤与脑囊虫病最易相互误诊。脑囊虫病误诊为脑瘤已见报道[1 ,2 ] ,脑瘤误诊为脑囊虫病报道甚少 [3] 。现将我院 1984～ 1998年间 9例脑瘤患者误诊为脑囊虫病报道如下。1　临床资料9例误诊为脑囊虫病的脑瘤 (胶质瘤和转移瘤 )患者均为1984～ 1998年在我院接受抗囊虫病原治疗的患者。 9例中男性6例 ,女性 3例 ,年龄 3…     

应用特异性单克隆抗体测定脑囊虫病患者血清及脑脊液中循环抗原

应用特异性单克隆抗体作DOT－ELISA和竞争ELISA检测65例脑囊虫病患者血清和脑脊液中游离循环Ag（FCAg）和循环免疫复合物抗原（CICAg）。血清FCAg阳性率分别为81．3％和81．5％（0．20～87．0μg／ml）,CICAg阳性率为95.4％和92．3％（0.19～120.0μg／ml）,脑脊液测定FCAg阳性单为78．5％和86.2％（0．32～25.0μg／ml）,CICAg阳性率为90.8％和93.8％（0.19～46.4μg／ml）。结果表明检测总循环抗原（包括游离部分和复合物部分）有利于提高脑囊虫病的诊断。     

Evaluation of Taenia solium and Taenia crassiceps cysticercal antigens for the serodiagnosis of neurocysticercosis

We evaluated the usefulness of seven cysticercal antigen extracts, four from Taenia solium cysticerci (whole parasite-TsoW, membrane-TsoMe, vesicular fluid-TsoVF and scolex-TsoSc) and three from T. crassiceps cysticerci (whole parasite-TcraW, membrane-TcraMe and vesicular fluid-TcraVF), for serodiagnosis of neurocysticercosis with an enzyme-linked immunosorbent assay (ELISA). Cysticercus-specific IgG were screened in serum samples from 23 patients with neurocysticercosis, 32 patients with other infections and 48 healthy persons. The best results were obtained with the TsoVF-ELISA (91.3% sensitivity; 96.2% specificity) and TcraVF-ELISA (91.3% sensitivity; 95% specificity). The ELISA done with whole parasite and membrane extracts from cysts of T. solium and T. crassiceps and the scolex extract from T. solium cysts showed a low performance in terms of sensitivity, ranging from 47.8% to 73.9%. None of the antigen preparations from T. solium and T. crassiceps cysticerci used in this study showed outstanding performance for the serodiagnosis of neurocysticercosis. However, considering the results obtained with the seven antigen preparations, vesicular fluid from T. solium and T. crassiceps cysticerci may be useful for detecting specific antibodies in sera from patients with neurocysticercosis.     

Recombinant expression of Taenia solium TS14 antigen and its utilization for immunodiagnosis of neurocysticercosis

In order to evaluate the potential use of TS14 antigen in an enzyme-linked immunosorbent assay (ELISA) for immunodiagnosis of neurocysticercosis (NC), its open reading frame (ORF) was amplified by RT-PCR from mRNA isolated from Taenia solium cysticerci. The ORF was subcloned into the expression vector pET-28a, and was used to transform Escherichia coli BL21 (DE3) cells to produce TS14 antigen. The His-tagged expressed protein was purified on a nickel affinity column. Using the _HISTS14 as antigen, ELISA was positive for 100% of cerebrospinal fluid (CSF) and 97% of serum samples from NC patients. No positive results were observed with sera and CSF samples from control groups. Cross-reactivity with sera from patients with schistosomiasis and Chagas’ disease was not observed. Serum samples from patients with taeniasis were evaluated and 2 of 13 cases showed reactivity in this assay. Our data indicate the usefulness of _HISTS14 in ELISA for an accurate and rapid assay for diagnosis of NC and seroepidemiological studies.     

Dot—ELISA在脑囊虫病诊断中的应用

本文报告用Dot-ELISA法检测脑囊虫病患者的血清和脑脊液的抗体。108例确诊为脑囊虫病患者的血清阳性率为81.6～96.1％,14例确诊为脑囊虫病患者脑脊液的阳性率为85.7～100％。54例正常人血清在1:40稀释度以上,均未出现假阳性;与并殖吸虫病患者及华支睾吸虫病患者血清均未出现交叉反应;与包虫病及脑血管疾病患者血清有一定的交叉反应。     

Evaluation of an IgG-ELISA strategy using Taenia solium metacestode somatic and excretory-secretory antigens for diagnosis of neurocysticercosis revealing biological stage of the larvae

Diagnosis of neurocysticercosis (NCC) is complicated because of the variability in clinical presentations and course of the disease where viability of parasite is a major determinant. The present study describes evaluation of ELISAs using Taenia solium metacestode somatic and excretory-secretory (ES) antigens for detection of anti-T. solium metacestode IgG antibodies in serum and cerebrospinal fluid (CSF). And results of the ELISAs in cases with a definitive diagnosis of NCC are correlated with the biological stages of the parasite such as live vesicular or degenerated stage. The sensitivity of the IgG-ELISA using ES antigen is observed to be much higher in serum (88.2%) than in CSF (64.28%) although it is only marginally higher in serum (76.4%) than in CSF (75%) when somatic antigen is used in the ELISA. Whereas, the specificities of the ELISA using either somatic or ES antigen for detection of IgG antibodies in serum (97.97%; 96.96%) and CSF (96.42%; 97.61%) are comparable. A strong association is observed between live stage of the parasite and detection of antibodies in sera and CSF from more number of NCC patients by ELISA using ES antigens. Similarly, detection of antibodies by ELISA using somatic antigens could be associated with the dead or degenerated stage of the parasite in brain. The IgG-ELISA strategy developed in the present study opens up an avenue for diagnosis of NCC in hospitals or in population prevalence studies. The use of crude extracts of ES proteins might improve the serodiagnosis of the cases of NCC carrying live vesicular stage of the parasite larvae.     

231例脑囊虫病患者脑脊液中循环抗原检测分析

本文报告应用抗猪囊尾蚴单克隆抗体４Ｆ８－ＥＬＩＳＡ对２３１例脑囊虫病患者脑脊液中循环抗原检测的结果。经与患者病史、临床分型和脑ＣＴ检查结果比较，发现脑脊液中ＣＡｇ与有无绦虫史无关。脑型合并皮下结节患者脑脊液中ＣＡｇ的阳性率明显高于单纯型脑囊虫病和皮下结节消失组的患者。ＣＡｇ检测的结果与囊尾蚴和脑部寄生的数量，病灶的新旧与部位等病理情况有关。表明ＭｃＡｂ（４Ｆ８）－ＥＬＩＳＡ不仅可用于脑囊虫病的诊断     

Detection of Cysticercus antigens and antibodies in cerebrospinal fluid of patients with chronic meningitis

Chronic meningitism is a less frequent manifestation of neurocysticercosis caused by Taenia solium cysticerci. In the present study we used Co-agglutination (Co-A), a simple and rapid slide agglutination test to detect specific Cysticercus antigen in the 67 cerebrospinal fluid (CSF) samples from patients with chronic meningitis of unknown etiology. The results were compared with that of ELISA for detection of antibodies. Among these samples four (5.97%) were positive for Cysticercus antigen by Co-A test and six (8.95%) were positive for antibodies by ELISA. Two samples were positive by both Co-A and ELISA, two were positive only by Co-A and four were positive only by ELISA. In the present study, although Cysticercus antigen and antibodies were present in CSF samples from eight (11.94%) patients, we cannot affirm that all the cases of chronic meningitis are due to cysticercosis, but for any case of chronic meningitis of unknown origin, it would be useful to consider the possibility of cysticercal meningitis.     

Excretory/secretory antigens (ES) from in-vitro cultures of Taenia crassiceps cysticerci,and use of an anti-ES monoclonal antibody for antigen detection in samples of cerebrospinal fluid from patients with neurocysticercosis

Antigens were obtained from cysticerci of the ORF strain of Taenia crassiceps, by culture of cysts in protein-free hybridoma medium (PFHM). Budding of new vesicles was observed after 24-48 h. Excretory/secretory (ES) antigens (peptides of <20 kDa) were recovered in the medium after culture for 48 h. SDS-PAGE analysis of vesicular-fluid (VF) antigens (obtained by rupturing T. crassiceps cysticerci in PFHM) and the ES antigens indicated partial homology between the two preparations. ES peptides of 18- and 14-kDa were recognized by polyclonal antibodies produced in rabbits immunized either with the VF antigens or with a total-antigen preparation of T. solium cysticerci. Antibodies present in samples of serum or cerebrospinal fluid (CSF) from patients with neurocysticercosis also reacted with ES peptides. An anti-ES monoclonal antibody detected antigens in the CSF from 10 patients with neurocysticercosis, showing the antigenic homology of the ES antigens with those of T. solium cysticerci in human infections.     

A rapid latex agglutination test for the detection of anti-cysticercus antibodies in cerebrospinal fluid (CSF)

Simple and rapid latex-based diagnostic tests have been used for detecting specific antigens or antibodies in several diseases. In this article, we present the preliminary results obtained with a latex agglutination test (LAT) for diagnosing neurocysticercosis by detection of antibodies in CSF. A total of 43 CSF samples were assayed by the LAT: 19 CSF samples from patients with neurocysticercosis and 24 CSF samples from patients with other neurologic disorders (neurosyphilis, n = 8; neurotoxoplasmosis, n = 3; viral meningitis, n = 4, chronic headache, n = 9). The LAT exhibited 89.5% sensitivity and 75% specificity. The use of LAT seems to be an additional approach for the screening of neurocysticercosis with advantage of simplicity and rapidity. Further studies could be performed using purified antigens and serum samples.