弓形虫抗血清IgG的制备及其检测效果研究

目的比较弓形虫不同免疫原所制备抗血清 IgG 的检测效果,优选高效价的抗体诊断试剂。方法制备弓形虫的细胞质抗原(C)、代谢抗原(S)。用抗原 C、C S、S 及活虫(P)各免疫2只家兔,收集抗血清经纯化而获得 IgG 抗体(多抗)并制备酶标记抗体;用上述4种多抗与对应的多抗-HRP 组合成4种双抗体夹心型 ELISA,检测人血清 CAg 和 C、S 抗原,评价试剂的敏感性;抗血清与疟原虫、隐孢子虫等抗原进行交叉反应性试验,评价试剂的特异性;采用捕获法检测人血清 IgM 抗体,比较4种多抗-HRP 的检测效果。结果各种 ELISA 夹心法同步检测 CAg 阳性标本7例,阴性样本8例,均未出现假阴性和假阳性反应,其 OD 均值的 P/N 比值依次为 C C 组33.5、S S 组27.8、CS CS 组21.2、P P 组13.2;C 和 S 抗原的最低检出量均为0.5μg/ml;与其它寄生虫未出现交叉反应。4种多抗-HRP 同步检测IgM 抗体阳性标本3例,阴性样本7例,均未出现假阴性和假阳性反应,其 OD 均值的 P/N 值最高的是抗 C-HRP。结论 4种抗体用于检测人血清 IgM 抗体、CAg、C 和 S 抗原以及其它抗原的结果表明,各种抗体试剂均具有良好的敏感性和特异性,其中以抗 C 抗体试剂最为满意。(2)4种组合形式的夹心ELISA,检测弓形虫抗原的差异无统计学意义,表明弓形虫 C 和 S 抗原间存在着共同抗原成分。     

弓形虫既往感染在孕期内对胎儿的影响

目的探讨既往感染者孕期内弓形虫的活动情况及对胎儿的影响。方法对68例抗弓形虫抗体IgG阳性、IgM阴性孕妇的血清、脐血采用酶联免疫吸附试验检测弓形虫抗体IgG、IgM、弓形虫循环抗原（CAg）,PCR法检测虫体DNA;胎盘样本采用直接涂片、匀浆涂片及PCR法观察弓形虫感染情况。结果68例弓形虫抗体IgG阳性孕妇中脐血弓形虫抗体IgG阳性28例,IgG胎盘垂直传播率41.2%;脐血弓形虫DNA阳性6例,宫内感染发生率8.8%;胎盘组织中虫体DNA阳性9例,宫内感染发生率13.2%。结论既往弓形虫感染孕期内仍可导致垂直传播。     

中枢神经系统弓形虫病的快速诊断方法的探讨

本文取ELISA检测弓形虫循环抗原（CAg）、特异性IgM及IgG抗体阳性者的脑脊液样本13例，作弓形虫的病原学对比检查，个别样本同时进行ELIB分析。结果，CAg与IgM双阳性2例、IgM阳性1例，采用涂片法查见弓形虫；CAg阳性2例、CAg与IgM双阳性1例，经接种分离而获得3株弓形虫；其余7例CAg阴性而IgM阳性或IgM与IgG双阳性的脑脊液样本，均卡检获病原体。1例CAg与病原学均为阳性的脑脊液样本，与弓形虫速落子代谢抗原同步进行ELIB，前者在50kD处出现明显的反应区带，后者分别在50、97和108kD处出现3条反应区带。从而表明，脑脊液中弓形虫CAg检出率与病原阳性率相符合，ELISA—CAg可作为中枢神经系统弓形虫病的快速诊断方法；而ELIB技术，对确定及区别弓形上现症患者，均具进一步探讨的价值。     

贵州省抑郁症患者刚地弓形虫感染状况调查及其基因型鉴定

目的了解贵州省抑郁症患者弓形虫感染状况及弓形虫基因型。方法 ELISA检测141例抑郁症患者和150例健康对照外周血弓形虫特异性抗体(IgG、IgM)和循环抗原(CAg),PCR扩增血样弓形虫高重复DNA片段(529 bp),多重-巢式PCR-限制性片段长度多态性(Mn-PCR-RFLP)方法鉴定弓形虫529 bp阳性样本的基因型。结果 ELISA检测结果显示,抑郁症患者组和健康对照组弓形虫抗体阳性率分别为21.3%(30/141)和7.3%(11/150),两者间差异有统计学意义(χ~2=11.674,P0.05);抑郁症患者组的IgG、IgM、CAg阳性率分别为18.4%(26/141)、1.4%(2/141)、1.4%(2/141);健康对照组IgG阳性率为7.3%(11/150),与抑郁症患者组间差异有统计学意义(P0.05),未检测到IgM和CAg。PCR结果显示,抑郁症患者组检测出1例弓形虫核酸阳性,经Mn-PCR-RFLP方法鉴定其基因型,结果为非典型Toxo DB#9(Chinese 1型)。结论贵州省抑郁症患者弓形虫抗体阳性率高于健康人群,1例抑郁症患者感染的弓形虫基因型为Chinese 1型。     

湘西地区小儿弓形虫感染检测结果调查分析

目的了解湘西自治州少数民族地区小儿人群弓形虫感染情况。方法选择1998年6月—2009年12月本院儿科住院患者529例,应用血清学酶联免疫吸附试验(ELISA)检测弓形虫的特异性抗体IgG、IgM和循环抗原(CAg)。结果 IgG阳性率10.02%,IgM阳性率8.32%,CAg阳性率0.95%,IgG+IgM阳性率11.53%,IgM+CAg阳性率1.13%,IgG+IgM+CAg阳性率为0.38%。与动物有密切接触史的患儿感染率高于无密切接触史的患儿(P0.05)。农村患儿感染率高于城市患儿(P0.05)。结论本地区儿科临床患者弓形虫感染总感染率为32.33%,发现多种临床患者都有感染现象,单纯IgG阳性有治疗价值。     

精神分裂症患者弓形虫感染状况的研究

目的 了解河北省张家口地区精神分裂症患者弓形虫感染状况,探讨弓形虫感染与精神分裂症之间的关系。方法 采用酶联免疫吸附试验对222例住院精神分裂症患者及78例健康体检者进行弓形虫循环抗原(CAg)、抗弓形虫抗体IgM、IgG检测。结果 张家口地区精神分裂症患者弓形虫感染率(14.86％)、CAg或IgM阳性率(10.36％)均高于健康体检者(3.85％)、(1.28％),两者之间差异均有显著性(P<0.05);33例弓形虫检测阳性患者中,CAg或IgM阳性率(69.7％)大于IgG阳性率(30.3％)(P<0.01)。结论 张家口地区精神分裂症患者弓形虫感染率高于健康人群,且活动性感染较严重,应加强对弓形虫病的认识,重视精神病患者的弓形虫感染情况。     

产妇弓形虫感染对新生儿影响的研究

目的 探讨孕妇弓形虫感染对胎儿的影响。方法 采用酶免三联诊断法对356例母婴杨对标本进行血清学检测。结果 产妇弓形虫感染率平均为19.1%,各单项检测分别为CAg15.2%、IgM2.5%、IgG3.4%;新生儿弓形虫感染率平均为17.7%,其脐血各单项检测分别为CAg16.9%、IgM0.3%、IgG2.2%。CAg垂直传播率为35.2%(19/54),IgG垂直传播率为33.3%(4/12)。在4例畸胎儿中,2例脐血弓形虫血清学阳性。结论 孕妇感染弓形虫有1/3机会可通过胎盘垂直感染胎儿,影响胎儿发育,重者致畸、致死。     

产妇弓形虫感染对新生儿影响的研究

目的探讨孕妇弓形虫感染对胎儿的影响.方法采用酶免三联诊断法对356例母婴配对标本进行血清学检测.结果产妇弓形虫感染率平均为1 9.1%,各单项检测分别为CAg 15.2%、IgM 2.5%、IgG 3.4%;新生儿弓形虫感染率平均为17.7%,其脐血各单项检测分别为CAg 1 6.9%、IgM 0.3%、IgG 2.2%.CAg垂直传播率为35.2%(19/54),IgG垂直传播率为33.3%(4/12).在4例畸胎儿中,2例脐血弓形虫血清学阳性.结论孕妇感染弓形虫有1/3机会可通过胎盘垂直感染胎儿,影响胎儿发育,重者致畸、致死.     

酶联免疫吸附试验检测弓形虫体IgA抗体及其临床应用的初步研究

试用弓形虫膜抗原ELISA对先天性弓形虫感染的畸形围产儿和后天获得性弓形虫感染者进行了特导性IgA抗体检测。11例先天性弓形虫宫内感染畸形围产儿的15份标本,6份检出IgA抗体,7份同时检出IgA;IgM抗体,两份仅检出IgM抗体。8份弓形虫抗体阳性的孕妇血清皆检出较强的IgA抗体。后天获得性弓形虫近期感染者32份血清,IgA抗体阳性3份,IgA、IgM抗体阳性8份,IgM抗体阳性21份;30份慢性感染者血清未有测到IgA抗体。对照组30份抗体阴性的孕妇血清及其围产儿的脐血检测结果抗体皆为阴性。IgA抗体是弓形虫感染早期的一个重要标志物,与IgM、IgG抗体同时检测,对临床急性弓形虫感染的诊断和疗效观察有着极其重大意义。     

弓形虫循环抗原诊断试剂的研究

目的　比较弓形虫不同免疫原所制备抗体检测CAg的效果 ,优选高效价的抗体诊断试剂。 方法　制备弓形虫的细胞质抗原 (C)、代谢抗原 (S)。用抗原C、C +S、S及活虫各免疫 2只家兔 ,收集抗血清经纯化而获得IgG抗体 (多抗 )并制备酶标记抗体 ;用上述 4种多抗与多抗 -HRP组合成 16种双抗体夹心型ELISA ,检测人血清CAg和C、S抗原 ,评价试剂的敏感性 ;抗血清与疟原虫、隐孢子虫等抗原进行交叉反应性试验 ,评价试剂的特异性。结果　各种ELISA夹心法同步检测CAg阳性样本 7例 ,阴性样本 8例 ,均未出现假阴性和假阳性反应 ,其OD均值的P/N比值依次为C +C组 33 5、S +S组 2 7 8、CS +CS组 2 1 2、P +P组 13 2 ;C和S抗原的最低检出量均为 0 5 μg/ml;与其它寄生虫未出现交叉反应。 结论　 (1) 4种抗体用于检测人血清CAg、C和S抗原以及其它抗原的结果表明 ,各种抗体试剂均具有良好的敏感性和特异性 ,其中以抗C抗体试剂最为满意。 (2 ) 16种组合形式的夹心ELISA ,检测弓形虫抗原的差异无统计学意义 ,表明弓形虫C和S抗原间存在着共同抗原成分。     

Kinetics of antibodies in sera, saliva, and urine samples from adult patients with primary or secondary dengue 3 virus infections.

OBJECTIVES: The kinetics of three serological markers (IgM, IgA, and IgG) in serum, saliva, and urine samples from adult patients with primary or secondary dengue infection were studied. DESIGN: Serum, saliva, and urine samples were collected from 22 patients with clinical and confirmed dengue 3 virus infection during the outbreak in Havana City in 2001. They were tested by capture IgM (MAC-ELISA), IgA (AAC-ELISA), and IgE (EAC-ELISA) and IgG ELISA inhibition method (EIM) to detect specific dengue antibodies. RESULTS: Similar kinetics were observed in IgM, IgA, and IgG antibodies in saliva and IgA and IgG in urine samples from secondary cases compared with kinetics in serum samples, although the values were lower. No IgG antibody was detected in saliva and urine samples in primary cases and IgM antibody was not detected in urine samples from either primary or secondary infection. All secondary cases were positive for IgG in saliva and urine samples at day 7. The kinetics of specific IgE antibodies in primary and secondary cases were different. CONCLUSIONS: The kinetics of three serological markers (IgM, IgA, and IgG) in serum, saliva, and urine samples from adult patients with primary or secondary dengue 3 virus infection were studied for the first time, showing its behavior and usefulness in dengue virus diagnosis. The specific IgE could play a role as a serological marker in secondary infections.     

ELISA检测妇科肿瘤患者伴发弓形虫感染的调查

[目的 ]了解妇科宫颈癌患者伴发弓形虫感染的情况。[方法 ]采用 EL ISA法检测弓形虫 Ig G、Ig M抗体和循环抗原。 [结果 ]宫颈癌、妇科良性肿瘤、其它疾患妇女 3组人群的弓形虫感染率分别为 :44 .0 % (2 2 / 5 0 )、2 5 .4% (15 / 5 9)和 2 0 .3% (2 8/ 138)。 3组人群 Ig G阳性率分别为 2 0 .0 0 %、11.86 %和 10 .87% ,Ig M阳性率分别为2 0 .0 0 %、13.5 6 %和 8.70 % ,循环抗原阳性率分别为 14.0 0 %、5 .0 9%和 1.45 %。 [结论 ]宫颈癌患者弓形虫感染率明显高于其它两组人群 ,以循环抗原和 Ig M阳性尤为明显     

棘球蚴病患者IgG抗体阴性反应血清再检测的研究

目的　探索棘球蚴病患者抗体应答假阴性反应原因 ,以改进棘球蚴病的免疫诊断方法。　方法　采用间接ELISA和双抗体夹心ELISA方法 ,检测 42例IgG抗体阴性反应棘球蚴病患者血清的IgG亚类 (IgG1、IgG2、IgG3和IgG4 )、IgA、IgM、IgE抗体及抗原和循环免疫复合物。　 结果　 42例阴性血清中 ,32例IgG亚类或IgA、IgM、IgE抗体阳性 ,1 0例血清抗体全部阴性。其中IgG1、IgG4及IgA、IgM、IgE的检出率明显高于正常人 ,分别为 42 .9%、1 1 .9%、2 8.6 %、2 6 .2 %和 2 1 .4 %。小儿的IgM高于成人。肝棘球蚴病患者的IgG亚类高于肺棘球蚴病患者。IgG1与其它抗体联合检测 ,以IgG1 +IgA +IgM检出率最高 ,为 64 .3 %。IgG阴性患者血清的CAg和CIC阳性率分别为 2 8.57%及30 .95 %。　结论　抗棘球蚴总IgG抗体表达水平低下 ,抗体表达种类不同及循环免疫复合物的形成 ,是造成棘球蚴病患者IgG抗体反应阴性的主要原因。IgG1 +IgA +IgM检测可提高棘球蚴病患者的诊断率     

Toxoplasma gondii: detection of antibodies in human saliva and serum

Saliva samples from 27 patients with a recent toxoplasma infection were tested for specific IgG, IgM and IgA antibodies to Toxoplasma gondii. Thirteen of the 27 saliva samples were positive for IgG anti-T. gondii by direct agglutination and 8 of the 27 were positive for IgM anti-T. gondii by an immunosorbent agglutination assay. Twenty of the 27 saliva samples were positive for IgG antibody on toxoplasma immunoblots with three major immunodominant antigens; 38, 30 and 35 kDa. IgA results on toxoplasma immunoblots were positive for all three groups tested, recently infected patients, chronically infected and seronegative adults without distinguishing between them. The 35 and 43 kDa antigens were the most frequently detected proteins. IgM in saliva gave negative or very weak reactions. None of the eight seronegative or the 17 chronically infected adults gave positive results in any of the tests performed to detect IgG or IgM in saliva. Serial saliva and serum samples from a laboratory-infected patient were collected and tested for toxoplasma-specific IgG, IgM and IgA. IgG in saliva was detected by 27 days post infection (p.i.) and was negative by 81 days p.i.; it detected mainly the 38 and 30 kDa antigens. IgM in saliva was detected by 11 days p.i. and was negative by 81 days p.i., with no reaction on immunoblots.     

Serodiagnosis of cryptosporidiosis in Italian HIV-positive patients by means of an oocyst soluble antigen in an ELISA.

A sensitive, specific, and reproducible ELISA which incorporated an oocyst soluble antigen was used in order to detect specific Cryptosporidium immunoglobulins G and M in Italian HIV-positive patients. The soluble antigen was prepared from purified oocysts collected from experimentally infected calves. The serum working dilution was established at 1 in 50. Of parasitologically positive and HIV-positive patients, 95% showed specific IgG or IgM or both in their serum. Of HIV-positive patients, some of whom had diarrhoea of uncertain aetiology, 15.8% were found to have specific IgG. Both specific IgG and IgM were detected in the serum of an HIV-positive patient 1 year before the shedding of oocysts in the faeces. Sixteen (5.3%) of 300 presumed healthy people, positive for specific IgG, were all IgM-negative. Any significant cross-reactions with other parasitic infections were not observed. With a serum dilution greater than 1 in 25, only a low degree of positivity was observed with samples highly IgG-positive for Toxoplasma species. The ELISA showed a higher than expected prevalence of Cryptosporidium infection in Italian HIV-positive patients.