ELISA和TRUST检测梅毒螺旋体方法比较分析

目的 比较ELISA(酶联免疫吸附试验)、TRUST(甲苯胺红不加热血清试验)法诊断梅毒螺旋体感染的方法学差异.方法 用目前国内最常用的诊断梅毒螺旋体感染的TRUST试剂及ELISA试剂检测722例标本,同时与TPPA(梅毒螺旋体明胶凝集试验)的检测结果进行比较,从而得到各试验的假阴性率和假阳性率.结果 对722份样本的检测中,ELISA和TPPA的阳性符合率为98.26％,假阴性率和假阳性率分别为0.46％和1.91％,2种方法差异无统计学意义(P ＞0.05); TRUST和TPPA的阳性符合率为84.39％,假阴性率和假阳性率分别为21.08％和5.10％;ELISA和TURST检出率差异有统计学意义(P＜0.01).结论 ELISA测定梅毒螺旋体的方法在日常大量标本检验时优于TRUST.     

ELISA与IHA测定弓形虫抗体评价及正确运用

<正> 用ELISA与IHA同时测定2252份人血清,IHA阳性率4.00％,ELISA阳性率4.57％,P>0.05。其中共同阳性89份,共同阴性2148份,ELISA阳性而IHA阴性14份,ELISA阴性而IHA阳性1份,阳性符合率85.58％,总符合率99.33％。对38份不同血凝滴度的标本,用ELISA稀释定量,1:16~+5例中,ELISA滴度均<1:200;1:16~(+++)5例中,有2例ELISA滴度1:400;1:64~(++)16例中,有1例ELISA为1:200,>1:64的12例中,ELISA滴度均>1:400,最高1:12800。IHA与ELISA的阳性误差往往出自结果在临界值附近的标本。抗人IgG-ELISA不适于动物与禽类血清抗体检测,使用IHA,能显示其优越性。本文测定结果见(附表)。     

应用生物素-亲和素系统检测弓形虫感染家兔特异性循环免疫复合物的实验研究

本文应用抗弓形虫单克隆抗体E_8株,亲和素和生物素标记的过氧化物酶复合物(ABC)为标记物的ABC-ELISA检测弓形虫感染家兔的特异性循环免疫复合物(CIC),并与常规ELISA、聚乙二醇沉淀试验(PEG)进行对比。 一、检测弓形虫感染兔(33只)及血吸虫感染兔(25只)血清CIC、ABC-ELISA检测CIC的阳性率为100％,PEG试验为96.9％:健康兔(29只)血清对照试验阴性符合率分别为100％及93.1％,但两法与血     

ELISA检测蛔虫感染者血清抗体的观察

采用ELISA检测蛔虫病流行区血清259例,其中193例粪检蛔虫卵阳性者,血清抗体的阳性符合率为93.8％:112例健康胎儿血清的阴性符合率为96.4％;56例肠道蠕虫卵阴性者血清的阴性率为82.1％;10例蛔虫卵阴性而肠道其它线虫卵阳性者血清的阳性率达30％。结果表明,此法诊断蛔虫感染具有较高的敏感性,但与肠道其它线虫感染血清有一定交叉反应。     

青海省果洛州人与动物棘球蚴病调查

目的通过对青海省果洛州人群与动物棘球蚴病的调查与分析,了解当地棘球蚴病在人群和动物中的流行与分布状况。方法应用B超对人群进行患病率调查;囊液抗原间接血凝试验(IHA)法筛查人群棘球蚴抗体的阳性率;并分别用重组Em18(rEm18)、重组抗原B(rAgB)和Eg囊液抗原(EgcF),对随机选取的B超检查和IHA检测阳性者进行ELISA试验,与B超诊断结果进行符合率比较,评价泡型棘球蚴病(AE)、囊型棘球蚴病(CE)的分布情况;应用ELISA检测家犬和藏狐粪抗原阳性率;现场检查牛、羊胸腹腔,确定牛、羊感染率。结果人群调查:①B超检查2826人,总患病率为9.45%。血清IHA法检查1113人,阳性率25.79%。女性人群的患病率和血清阳性率均显著高于男性;②血清囊液抗原IHA检测阳性者(108)与ELISA法检测EgcF,阳性符合率为66.94%;3种重组抗原检测,ELISA法与IHA法阳性符合率为75.00%。③B超法诊断棘球蚴病(88人)与ELISA法检测,符合率EgcF为94.32%,rAgB为68.18%,rEm18为5l-4%。EgcF阳性率符合率为69.44%;3种抗原检测与IHA检测总体阳性符合率为75.00%。61例CE患者中,血清ELISA法检测,EgcF阳性率为96.72%,rAgB为68.85%,rEm18为33.33%;26例AE患者中,EgcF和rEm18阳性率均为92.31%,rAgB为65.39%。动物调查:①高原鼠兔棘球蚴感染率21.74%。经cytb基因分析,证实有多房棘球绦虫(E.multilocularis)和石渠棘球绦虫(E.shiquicus)感染。绵羊棘球蚴感染率82.61%,牦牛棘球蚴感染率78.52%。②野犬、藏狐、猞猁均有不同程度的棘球绦虫感染。结论青海省果洛州人群与动物中存在棘球蚴病的高度流行。E.shiquicus种的幼虫和成虫分别在高原鼠兔和藏狐体内寄生已得到证实,提示果洛州为3种棘球蚴的混合流行区。     

华支睾吸虫重组半胱氨酸蛋白酶用于血清学调查的效果评价

目的 以华支睾吸虫重组半胱氨酸蛋白酶(CsCp)为ELISA包被抗原检测流行人群血清特异性抗体水平,与成虫粗抗原比较,评价其用于华支睾吸虫病血清学调查的价值. 方法 以佛山市南海区的1个行政村为调查点,用改良加藤厚涂片法检查粪便华支睾吸虫虫卵.同时采集血清,分别以rCsCP与成虫粗抗原(CsCAA)的ELISA方法检测血清特异性抗体水平,对粪检及两种抗原的ELISA结果进行统计学分析. 结果 接受血清检测的97人中,华支睾吸虫虫卵阳性率60.8%,CsCAA-ELISA阳性率75.3%,rCsCP-ELISA阳性率76.3%,特异性抗体阳性率高于虫卵阳性率(P<0.05),不同抗原ELISA阳性率差异无统计学意义(P>0.05);CsCAA-ELISA、rCsCP- ELISA结果与粪检结果的符合率分别为56.7%和55.7%,两种抗原ELISA结果的符合率为82.5%.59例粪检阳性者CsCAA-ELISA和rCsCP-ELISA阳性率均为76.3%,38例粪检阴性者CsCAA-ELISA和rCsCP-ELISA阳性率分别为73.7%和76.3%. 结论 以rCsCP为包被抗原,用ELISA检测华支睾吸虫病流行区人群血清特异性IgG抗体与使用CsCAA的检测结果有较高的符合率,但同样不能区分现症与既往感染.     

McAb检测囊尾蚴循环抗原诊断脑囊虫病试剂盒的研制

本文报道了McAb(4F_2)ELISA检测囊尾蚴循环抗原诊断脑囊虫病试剂盒的研制。实验对227例脑囊虫病患者脑脊液进行检测,循环抗原阳性率为84.58％,其中活动型脑囊虫病患者171例,循环抗原阳性率为93.57％。56例非活动型脑囊虫病患者,循环抗原阳性率为57.14％,前者的循环抗原强度也明显高于后者。83例患有其它中枢神经系统疾病的患者,1例阳性,阴性符合率为98.80％。本试验方法简便,结果客观,对脑囊虫病特别是活动型脑囊虫病敏感性高、特异性强。     

金免疫斑点渗滤法检测日本血吸虫抗体的应用

建立了检测血吸虫病人血清抗体的金免疫斑点渗滤法（DIGFA）。以该法检测107例粪检阳性血吸虫病人血清,阳性率为96．2％,119份健康人血清均为阴性;与常规ELISA对照,符合率达99．1％,显示两法敏感性和特异性相近,但DIGFA操作更为简便快速。     

金免疫斑点渗滤法检测日本血吸虫抗体的应用

建立了检测血吸虫病人血清抗体的金免疫斑点渗滤法(DIGFA)。以该法检测107份粪检阳性血吸虫病人血清,阳性率为96.2%。119份健康人血清均为阴性。与常规ELISA对照,符合率达99.1%。显示两法敏感性和特异性相近,但DIGFA操作更为简便快速。     

蛋白芯片法检测TORCH-IgM抗体的临床应用研究

目的评价TORCH蛋白芯片诊断致畸病原体的临床应用价值。方法采用TORCH蛋白芯片法与ELISA法对230份孕妇血清标本作平行检测;用蛋白芯片法对卫生部临床检验中心室问质评样本进行检测。结果TORCH蛋白芯片法与ELISA法各项指标检测结果均具有较好的一致性（P〉0.05）,其中TOX的阴、阳性符合率均为100％;RV的阳性符合率为85.7％,阴性符合率为99.6％;CMV的阳性符合率为91.7％,阴性符合率为99.1％;Hsv-Ⅱ的阳性符合率为93.8％,阴性符合率为99.1％。室间质评样本的检测结果与卫生部临床检验中心公布结果一致。结论蛋白芯片法检测TORCH-IgM抗体具有简便、快速,敏感性较高和特异性强等优点,是临床优生优育辅助诊断的有效方法,值得推广使用。     

COMPARISON OF ELISA AND lEA FOR ESTIMATION OF ANTIBODY LEVELS OF CATTLE TO THEILERIA ANNULATA VACCINE

Bovine tropical Theileriosis caused by Theileria annulata is an economically important disease of cattle. An enzyme linked immunosorbent assay (ELISA) was used to determine antibody levels in vaccinated and unvaccinated cattle, using cellular schizont as antigen and its results were compared with immunofluorescent assay (IFA). For this test 126 sera collected (105 vaccinated, 31 not vaccinated) from cows and assayed with ELISA which among them 104 sera were positive and 32 sera were negative. Same sample assayed with IFA in which 99 were positive sera and 37 were negative sera. Thereby the sensitivity and specificity of this ELISA on comparsion with lEA were 95.5% and 66.6% respectively. This study revealed that ELISA could be successfully used for both differentiating vaccinated and not vaccinated cattle and obtaining the titer of vaccinated cattle.     

Diagnosis of Fasciola gigantica infection using a monoclonal antibody-based sandwich ELISA for detection of circulating cathepsin B3 protease

A reliable monoclonal antibody (MoAb)-based sandwich enzyme-linked immunosorbent assay (sandwich ELISA) was developed for the detection of circulating cathepsin B3 protease (CatB3) in the sera from mice experimentally infected with Fasciola gigantica and cattle naturally infected with the same parasite. The MoAb 2F9 and biotinylated rabbit polyclonal anti-recombinant CatB3 antibody were selected due to their high reactivities and specificities to F. gigantica CatB3 antigen based on indirect ELISA and immunoblotting. The lower detection limit of the sandwich ELISA assay was 10, 100 and 400 pg/ml, when applied for the detection of rCatB3 antigen and CatB3 in whole body (WB) of newly excysted juveniles (NEJ) and metacercariae (Met) of F. gigantica, respectively. This sandwich ELISA assay could detect F. gigantica infection from day 1 to 35 post infection and revealed that circulating level of CatB3 peaked at day 1 post infection. In contrast, the antibody detection by indirect ELISA could only demonstrate the antibody level from 35 days post infection. The reliability of the assay method was evaluated using serum samples from mice infected with F. gigantica or Schistosoma mansoni, and hamsters infected with Opisthorchis viverrini, as well as normal mice and hamsters. In addition, sera from cattle infected with Paramphistomum cervi, Strongylid, Trichuris sp. and Strongyloides sp., as well as sera from normal cattle were also assessed. In experimental mice, the diagnostic sensitivity, specificity, positive predictive value, negative predictive value, false positive rate, false negative rate and accuracy of ELISA were 95%, 100%, 100%, 97.9%, 0%, 5.3% and 98.5%, while in natural cattle they were 96.7%, 100%, 100%, 98.5%, 0%, 3.4% and 98.9%, respectively. Hence, this assay method showed high efficient and precision for early diagnosis of fasciolosis by F. gigantica.     

Value of ELISA using A60 antigen in the diagnosis of active pulmonary tuberculosis

This investigation was undertaken to assess the effectiveness of an enzyme-linked immunosorbent assay (ELISA) using A60 antigen in ascertaining diagnosis in hospitalized patients suspected to have pulmonary tuberculosis (TB) but with negative sputum stains. Cultures were performed to confirm active or inactive disease. IgG and IgM antibody activity was determined by adding a 1:100 dilution of serum to plates coated with A60 antigen. After addition of peroxidase-conjugated antihuman IgG or IgM and color development, optical density (OD) was determined. A total of 83 patients was studied, taking into account their current disease status and prior history. Using as a cutoff value the mean value +/- 2 SD measured in the negative culture, no TB history group, that is, OD = 0.50 for IgG measurements and 0.43 for IgM measurements, the sensitivity, specificity, and positive predictive value of IgG measurements were equal to 48, 71, and 50%, respectively. Using IgM measurements, these parameters were equal to 76, 98, and 95%, respectively. Combining the results of IgG and IgM measurements, sensitivity, specificity, and positive predictive value were equal to 68, 100, and 100%, respectively. Thus, the ELISA described here can greatly facilitate the diagnosis of TB in patients with negative smears.     

Diagnosis of intestinal amebiasis using salivary IgA antibody detection

Attempts were made to use soluble antigen extract of strain HK-9 of Entamoeba histolytica to detect salivary IgA antibodies in intestinal amebiasis patients by using ELISA. Total salivary samples of 109 individuals were divided into four groups. Group I comprised 32 patients whose stools were positive only for E. histolytica cysts and/or trophozoites. Group II comprised 12 individuals whose stools were positive for E. histolytica and other intestinal parasites. Group III comprised 36 individuals whose stools were negative for E. histolytica but contained other intestinal parasites such as E. coli, E. nana, Blastocystis hominis, Trichomonas hominis, Giardia lamblia, Opisthorchis viverrini, and hookworm. Group IV comprised 29 healthy individuals whose stools were free from any intestinal parasitic infections. Based on the mean optical density, OD + 2SD of the results from 29 parasitologically negative healthy individuals, the cut-off OD value for salivary IgA antibodies was 1.265. Therefore, the assays were positive in 14 out of 32 (43.75%) of group I and 2 out of 12 (16.6%) of group II. The assays were positive in 16 out of 36 (44.44%) for group III whereas 2 out of 29 (6.90%) for group IV were positive. The overall sensitivity and specificity of the assays were 36% and 72%, respectively. The false positive rate was 28% and the false negative rate was 64%. The predictive values of positive and negative results were 47% and 63%, respectively. The diagnostic accuracy of ELISA for the presence of salivary IgA antibodies was 58%.     

河南省信阳市动物中新布尼亚病毒感染情况调查

目的 了解河南省信阳市动物中新布尼亚病毒感染情况,为研究其传播途径和防治策略奠定基础。方法 在信阳市商城县和光山县采集动物血液,应用荧光RT-PCR方法检测血液标本中是否携带新布尼亚病毒核酸,采用双抗原夹心ELISA方法检测新布尼亚病毒总抗体。结果 在调查点采集到5种动物血清标本合计312份,结果均未检测到新布尼亚病毒核酸;141份动物血清检测到新布尼亚病毒总抗体,总体阳性率为45.19%,鼠、牛、羊、猪和狗分别为1.06%,100.00%、76.27%、3.57%和75.00%,5种动物的抗体阳性率有统计学意义。结论 河南省信阳市牛、羊、狗等家养动物中存在新布尼亚病毒抗体,可能为该病毒的储存宿主。     

ABC-ELISA检测喜马拉雅旱獭鼠疫FI抗体的研究

用ABC-ELISA和常规ELISA检测了19份血凝阳性喜马拉雅旱獭(Marmota himalayana)血清和48份血凝阴性喜马拉雅旱獭血清及247份近年采自疫区的该旱獭血清,比较了两法的敏感性与特异性。两法的阳性、阴性符合率均为100％,但检测阳性血清的OD均值ABC-ELISA法明显地比常规ELISA法高(P<0.01)。ABC-ELISA与常规ELISA法对疫区血清标本的阳性检出率分别为9.7％与8.1%,后者出现1.6％的漏检率,滴度也显著低于前者(P<0.01)。表明ABC-ELISA用于检测旱獭鼠疫FI抗体可明显提高ELISA法的敏感性,且能保持良好的特异性。     

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。     

ELISA、胶体硒和免疫试鸭验检测HIV抗体结果分析

目的 复检和确认初筛HIV抗体阳性血清，并对比分析试验结果。方法 对初筛阳性血清，用胶体硒和ELISA试验复检，任一复检试验阳性的血清再用westernblot（WB）试验确认。结果 224份初筛阳性血清经复检84份阳性，WB确认61份HIV—1抗体阳性，不确定10份，阴性13份。以WB结果为标准，ELISA、胶体硒法的阳性符合率分别为84．72％和82．43％。ELISA的敏感性为100％，特异性为96．84％，胶体硒法的敏感性为100％，特异性为95．03％。阳性、不确定和阴性组血清EI，ISA平均S／CO分别为20．550，2．244和1．434。ELISA和胶体硒法复检同时阳性，在阳性组、不确定组、阴性组分别为100％（61／61），10％（1／10）和0（0／13）。结论 胶体硒和ELISA复检可排除大部分初筛假阳性，但两种复检试验均存在一定的假阳性。3种试验的不符情况仅出现在HIV抗体阴性和不确定组，确定HIV感染依赖于WB确认试验。     

PCR-ELISA检测大鼠卡氏肺孢子虫DNA的研究

目的　建立卡氏肺孢子虫 (P.c )DNA的聚合酶链反应-酶联免疫吸附测定(PCR-ELISA)方法 ,并探讨其应用价值。　方法　实验组患卡氏肺孢子虫肺炎的SD大鼠和Wistar大鼠各 2 8只 ,采用PCR法扩增大鼠肺组织DNA和支气管肺泡灌洗液 (BALF)DNA ,用PCR ELISA检测其扩增产物。 2 8只患病大鼠分别制作肺组织印片及BALF涂片 ,姬姆萨 (Giemsa)染色镜检 10 0个视野中有无P .c包囊 (或滋养体 ) ,与PCR ELISA检测扩增产物结果比较。　结果　两种方法检测大鼠肺组织DNA阳性率及BALFDNA阳性率 ,结果相同 ,均分别为 96.4% (27/28)和100% (28/28)。Giemsa染色镜检P.c包囊 (或滋养体 ) ,结果为阳性的大鼠 ,PCR ELISA检测扩增产物结果也均为阳性。阴性对照组 ,两种大鼠的肺组织和BALF各10份标本 ,均有1只大鼠阳性。　结论　PCR ELISA检测大鼠卡氏肺孢子虫DNA ,敏感性较高 ,特异性较好 ,操作简便 ,具有实用价值。     

Evaluation of brucellin skin test for bovine brucellosis

The diagnostic efficiency of the Brucellin skin test for bovine brucellosis was evaluated using cattle of known history. The test was negative in six out of 14 heifers (42.9%) infected with virulent Brucella abortus (Br. abortus) strain 544. In four cattle vaccinated with a reduced dose of Br. abortus strain 19 vaccine (5 X 10(9) live organisms) the skin test became positive in all the animals but two weeks after immunization. However, all the vaccinates became negative 14 weeks after vaccination, whereas nine out of 14 heifers (64.3%) vaccinated with killed Br. abortus 45/20 adjuvant vaccine (Duphavac vaccine) were still positive 18 months post immunization. Four control cattle were persistently negative. It was considered that the procedure would be most useful for testing non-exposed cattle and should be used as a screen test. It is not useful as a diagnostic test for cattle immunized with Duphavac vaccine. A recommendation is made for interpreting the Brucellin skin test for the diagnosis of bovine brucellosis in non-exposed herds.