5种市售弓形虫抗体检测试剂盒的评价

目的　评价国内公司研制的 5种弓形虫抗体检测试剂盒。方法　以敏感性、特异性、符合率和Youden指数作分析指标 ,比较 5种试剂盒对国外进口的 2种弓形虫抗体试剂盒筛选出的弓形虫IgG或IgM阳性血清和正常人血清的检测结果。结果　A、B、C 3种IgM试剂盒的检测敏感性和特异性分别为 0和 92 1%、6 7%和 10 0 %、以及 13 3%和 85 7% ,Youden指数分别为 - 0 0 8、0 0 7和 - 0 0 1,基本无诊断价值。A’、B’ 2种IgG试剂盒的检测敏感性分别为 96 4 %和 32 1% ,特异性为 98 4 %和 96 8% ,Youden指数为 0 95和 0 2 9。A’试剂盒与进口IgG试剂盒的符合率达到 97 8% ,诊断价值较大 ,而B’试剂盒与进口试剂盒的符合率仅为 76 9% ,A’、B’ 2种试剂盒之间的符合率也只有 76 9%。结论　国内研制的市售弓形虫试剂盒的检测效果与质量存在一定的问题 ,尤以IgM试剂盒为甚。     

两种抗原检测弓形虫病IgM的效果

目的观察弓形虫P30重组抗原及可溶性抗原检测弓形虫病IgM的效果。方法建立斑点免疫金渗滤法（DIGFA）。采用2种抗原分别包被在两条硝酸纤维素膜（NC）上,与待检血清中的相应IgM抗体结合,通过金标记兔抗人IgM直接显色,以检测弓形虫病IgM。2种抗原的DIGFA分别检测弓形虫IgM抗体阳性、类风湿、支原体肺炎、血吸虫病人和正常人血清。结果两种抗原建立的DIGFA检测试剂共检测5种血清样本166份,其中弓形虫感染IgM抗体阳性病人血清38份,弓形虫P30DIGFA敏感性为92.1%（35/38）,可溶性抗原DIGFA敏感性为81.6%（31/38）,差异无统计学意义（P〉0.05）。正常人群对照血清63份,检测全部阴性,特异性为100%。检测类风湿病人血清30份、血吸虫病人血清25份及支原体肺炎病人血清10份,仅类风湿病人血清出现阳性交叉反应,其中弓形虫P30DIGFA阳性2例,可溶性抗原DIGFA阳性3例。结论弓形虫P30重组抗原检测弓形虫病人血清IgM的DIGFA试剂可用于临床早期或急性期弓形虫病的诊断。     

3种方法检测弓形虫IgG抗体的效能

目的 目的 比较3种不同的检验方法对弓形虫IgG抗体的检测效能, 并进行分析。方法 方法 采用临床上常用的金标法、 间接血凝试验 （IHA）、 酶联免疫吸附实验（ELISA）, 对304份血清标本进行弓形虫IgG抗体平行检测, 比较各检测方法的敏 感性、 特异性和Youden指数。结果 结果 金标法、 IHA、 ELISA这3种方法对弓形虫IgG抗体检测的敏感性分别为85.5%、 89.8% 和91.9%, 三者之间差异没有统计学意义 （χ2 = 4.12, P > 0.05）; 特异性分别是92.4%、 96.6%和97.5%, 差异亦无统计学意义 （χ2 =4.06, P > 0.05）。ELISA法的检测效率和Youden指数分别为94.1%和0.89, 高于IHA法和金标法。结论 结论 ELISA法检 测弓形虫IgG抗体的敏感性和特异性较高, 可自动化, 适合于大规模的弓形虫IgG抗体筛查。     

血吸虫感染小鼠早期诊断抗原的研究

目的对已知的6种日本血吸虫抗原的早期诊断价值进行评价,为研制用于哨鼠早期诊断的免疫试剂提供候选抗原。方法用日本血吸虫尾蚴感染小鼠,收集感染前及感染后不同时间的小鼠血清。采用重组日本血吸虫中国大陆株23kDa膜蛋白大亲水性肽段与谷胱甘肽的融合表达蛋白（GST-HD）、可溶性虫卵抗原（SEA）、血吸虫四跨膜蛋白第二亲水基团（TSP2HD）、血吸虫虫卵蛋白（IPSE）、日本血吸虫虫卵毛蚴抗原（SjMP-10）及日本血吸虫信号蛋白（Sj14-3-3）作为诊断抗原,采用酶联免疫吸附试验检测感染血吸虫后不同时间小鼠血清中特异性抗体IgM和IgG水平,通过分析感染后不同时间点抗原特异性抗体水平变化及阳性率,筛选具有血吸虫感染早期诊断价值的抗原分子。采用免疫印迹试验进一步验证其对血吸虫急性感染早期诊断的价值。结果感染后第18、21、28天,抗GST-HD抗体IgM阳性率分别为60%、70%、100%,特异性IgG阳性率分别为40%、60%、90%;抗SEA抗体IgM阳性率为50%、60%、90%,特异性IgG阳性率为20%、50%、70%;抗TSP2HD抗体IgM阳性率为30%、40%、50%,特异性IgG阳性率为20%、30%、70%;抗IPSE抗体IgM阳性率为20%、30%、50%,特异性IgG阳性率为20%、30%、60%;抗SjMP-10抗体IgM阳性率为10%、20%、20%,特异性IgG阳性率为10%、20%、30%;抗Sj14-3-3抗体IgM阳性率为0、10%、20%,特异性IgG阳性率为0、10%、30%。以GST-HD融合蛋白和SEA为抗原,检测小鼠早期感染血吸虫的敏感性高于Sj14-3-3、IPSE、TSP、MP-10,检测IgM的敏感性高于IgG。免疫印迹试验结果显示,SEA中分子量在73kDa左右的蛋白条带可被感染后1周小鼠血清所识别,并随着时间推移反应加强。GST-HD最早出现反应的血清是感染后第10天小鼠血清,反应强度在感染后第5周达到最强。结论重组GST-HD融合蛋白与SEA中分子量约73kDa的蛋白分子具有血吸虫感染早期诊断价值,免疫印迹试验的敏感性比酶联免疫吸附试验高。     

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。     

人唾液、尿液中弓形虫抗原抗体的检测

目的探讨用唾液、尿液替代血清进行弓形虫感染免疫学诊断的可行性。方法用ELISA法对血清进行弓形虫循环抗原(CAg)、IgM、IgG抗体的检测,取上述各项免疫学指标阳性的血清样本80例,并随机抽取3项指标均为阴性的血清样本102例,分别与其唾液和尿液标本在同一反应板上同步进行测定,比较其阳性和阴性符合率。结果(1)554份血清样本中,CAg、IgM和IgG3项的阳性率分别为4.51%(25/554)、3.61%(20/554)、6.32%(35/554)。(2)唾液中检出IgM抗体阳性16例,与血清的阳性符合率为80%(16/20),检测CAg、IgG抗体均为阴性。(3)检测尿液中的抗原抗体均无一例阳性。结论采用唾液标本检测弓形虫IgM抗体,在弓形虫感染的诊断及流行病学调查中均具有一定的应用前景。     

乙酰螺旋霉素联合阿奇霉素治疗孕期弓形虫感染的临床效果

目的 目的 探讨螺旋霉素联合阿奇霉素治疗孕期弓形虫感染的综合治疗方法。方法 方法 应用ELISA方法检测孕妇血清 特异性弓形虫IgG和IgM抗体, 并采用PCR方法检测羊水中弓形虫感染情况; 选取其中3例弓形虫IgM抗体阳性孕妇作为 治疗对象, 应用螺旋霉素联合阿奇霉素进行综合治疗, 通过血清学特异性抗体检测和PCR检测结果初步判断其疗效。结 结 果 果 经ELISA检测, 孕期妇女血清特异性弓形虫IgG和IgM抗体的阳性率分别为5.97% （17/285） 和1.05% （3/285）, 对3例血 清IgM抗体阳性孕妇的羊水进行PCR检测特异性弓形虫基因, 有2例呈阳性。经过2种药物的联合治疗, 这2例孕妇血清 特异性弓形虫IgM抗体滴度阴转, PCR检测产妇脐带血中弓形虫基因为阴性。结论 结论 螺旋霉素联合阿奇霉素治疗孕期弓 形虫感染安全、 有效。     

金标渗滤法检测疟疾抗体的研究

目的建立一种适用于现场使用的疟疾抗体快速检测方法。方法以恶性疟原虫可溶性抗原作为包被抗原,胶体金标记羊抗人IgG为第二抗体,建立快速检测疟疾抗体的金标渗滤法(DIGFA)。采用双盲法,与间接荧光法(IFAT)平行检测23份恶性疟和45份间日疟患者血清以及101份健康对照者血清,比较敏感性、特异性、Youden指数以及其他试验条件,以评价检测效果。结果DIGFA对恶性疟和间日疟的检测敏感性分别为95.65%和71.11%,对健康对照者的特异性为97.03%,Youden指数分别为0.93和0.68;IFAT的敏感性分别为95.65%和84.44%,特异性为96.04%,Youden指数分别为0.92和0.80。两法总Youden指数分别为0.76和0.84,总符合率为90.48%。结论DIGFA操作简易,适用于现场疟疾抗体的检测和监测。     

斑点免疫金渗滤法检测弓形虫感染效果

目的建立一种简捷、准确的弓形虫感染检测方法。方法将弓形虫P30抗原包被在硝酸纤维素膜上,与待检血清中的相应抗体结合,通过金标记Protein-A蛋白直接显色。结果用该试剂检测5种血清样本共221份,其中弓形虫感染病人血清58份,敏感性为96.55%(56/58)。正常人群对照血清68份,检测结果全部为阴性,特异性为100%。检测25份类风湿病人血清、18份血吸虫病人血清及22份肺炎支原体病人血清,仅有1例类风湿病人血清出现阳性交叉反应。结论用弓形虫P30为抗原,以金标记Protein-A蛋白为探针显色系统的斑点免疫金渗滤法,对检测弓形虫病人血清抗体具有较高的敏感性和特异性,且操作简单、快速。     

IEST检测感染犬弓首线虫幼虫鼠血清抗体的研究

用感染犬弓首线虫幼虫鼠肝石蜡切片作为抗原片进行免疫酶染色试验（IEST），检测30份犬弓首线虫幼虫感染鼠血清抗体（IgG），阳性率为96.67％。12份旋毛虫感染鼠血清及12份健康鼠血清抗体均为阴性，结果表明本法的敏感性和特异性均较高。     

检测弓形虫IgM抗体的3种试剂盒的比较分析

目的 评价用于检测弓形虫IgM抗体的3种市售的试剂盒。方法 用3种试剂盒检测125份孕妇的血清,比较各试剂盒的阳性检出率、敏感性、特异性和试验效率等。结果 125份血清经Bi、Di和So试剂盒检测,分别有12．8％、15．2％和12．0％阳性,3种试剂盒检测血清的阳性率之间无显著性差异（P＞0．05）。Bi、Di和So试剂的敏感性和特异性分别为88．9％与100％、100％与99．1％和83．3％与100％,试验效率分别为98．4％、99．2％和97．6％。用WHO国际生物标准化实验室的抗号形虫血清标准品定量测得3种试剂盒的灵敏度都是0．31IU／ml。Bi试剂盒与Di试剂盒的总符合率为97．6％;Di试剂盒与So试剂盒的总符合率为96．8％;Bi试剂盒与So试剂盒的总符合率为97．6％。结论 这3种试剂盒都能用于弓形虫IgM抗体的常规普筛和血清学诊断。     

Toxoplasma gondii infection among chronic hepatitis C patients:A casecontrol study

Objective:To determine the detection rate of anti-Toxoplasma gondii(T.gondii) IgG and IgM in chronic HCV patients attending the Department of Tropical Medicine Mansoura University hospital in Egypt.Methods:This study included 120 adult chronic HCV patients.81 decompensate cirrhosis(late-stage)and 39 chronic HCV non cirrhotic patients(early-stage) and40 healthy blood donors as controls.Serum samples uere examined for anti-Toxoplasma IgM and anti-Toxoplasma IgG antibodies by ELISA.Real-time RT-polymerase chain reaction assay was done for quantitation of hepatitis C virus.Results:Anti-T.gondii IgG antibodies were detected in 75(92.6%) of 81 late-stage cirrhotic patients.30(76.9%) of the 39 chronic HCV non cirrhotic patients(early-Stage) and in 6(15ft) of 40 controls with statistically significant difference(P0.001).Anti-T.gondii IgM antibodies were found in 11(13.6%) in late stage patients,5(12.8%)in early stage and in 3(7.5%) of controls with no statistical significant difference(P=0.610).There was no correlation between stage of fibrosis and IgM or IgG antibodies positivity in our studied groups(P=0.526).High IgG levels significantly correlated with high viral load(P=0.026).Conclusions:Our findings suggest that the serious opportunistic T.gondii infection represent a potential significant risk for chronic HCV patients.So.toxoplasmosis should be considered in their investigations and follow-up.     

Seroepidemiology of Toxoplasma gondii infection in Tepehuanos in Durango, Mexico

The epidemiology of Toxoplasma gondii infection is known to depend on a number of behavioral factors, including contact with animals and eating habits. The epidemiology in specific populations (i.e., in indigenous ethnic groups) is not well studied. We therefore investigated the presence of anti-Toxoplasma IgG and IgM antibodies in 156 Tepehuanos, an indigenous ethnic group in Durango State in northern Mexico, using ELISAs. Of the 156 Tepehuanos (mean age 31.03±16.71 years), 35 (22.4%) had anti-Toxoplasma IgG antibodies. IgG levels of 14-99, 100-150, and >150?IU/mL were found in 15 (42.9%), 1 (2.8%), and 19 (54.3%) anti-Toxoplasma IgG-positive Tepehuanos, respectively. Also, 15 (9.6%) subjects had anti-Toxoplasma IgM antibodies. Male Tepehuanos aged 31-50 years showed a significantly higher frequency of exposure to T. gondii than female Tepehuanos of the same age. Seroprevalence of anti-T. gondii IgM antibodies was significantly higher in Tepehuanos without education than those with 7-12 years of education. Tepehuanos suffering from frequent headaches had significantly higher seroprevalences of anti-T. gondii IgG and IgM antibodies than those without this clinical feature. In addition, Tepehuanos suffering from hearing impairment had significantly higher seroprevalence of anti-T. gondii IgM antibodies than those without this clinical feature. Logistic regression analysis did not show an association of T. gondii exposure with any behavioral characteristics, including contact with cats or eating habits. The seroprevalence of T. gondii exposure in Tepehuanos is comparable to that reported in other populations in North America; however, seroprevalence in the largely rural Tepehuanos is higher than that reported in a nearby urban center. Toxoplasma may be impacting the health of Tepehuanos. This is the first report of T. gondii infection in Tepehuanos, and the results should prove useful for the design of preventive measures.     

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。     

Seroprevalence of Toxoplasma gondii, rubella and cytomegalovirus among pregnant women in southern Turkey

Primary infections caused by Toxoplasma gondii, rubella and cytomegalovirus (CMV) can lead to serious complications in pregnant women. The aim of this study was to determine the seroprevalence of Toxoplasma, rubella and CMV infections through antenatal screening. In this study, the consecutive records of 1652 pregnant women examined between the period March 2004 to January 2006 were included. The results of the antenatal screening for Toxoplasma, rubella and CMV during the first trimester of pregnancy were evaluated. Anti-Toxoplasma, anti-rubella and anti-CMV IgG and IgM antibodies were assayed using an enzyme linked immunosorbent assay method. Of the 1652 pregnant women tested, anti-Toxoplasma IgG antibody was found in 860 (52.1%) of the cases, while 9 (0.54%) of the subjects tested positive for anti-Toxoplasma IgM. Anti-rubella IgG and IgM antibodies were reactive in 1570 (95.0%), and in 9 (0.54%) of the tested women, respectively. Moreover, 1568 (94.9%) of them were found to be positive for anti-CMV IgG, while 7 (0.4%) tested positive for anti-CMV IgM. Consequently, because of the high seropositivity of T. gondii, rubella and CMV in the pregnant women, the country's health authorities should be alerted, and preventive measures should be taken.     

孕妇妊娠各期弓形虫感染与垂直传播的临床研究

目的研究孕妇妊娠各期弓形虫(TOX)感染及其垂直传播状况。方法采用酶联免疫捕获法检测孕妇及胎儿的流产组织或脐血TOX-IgM抗体。结果检测5 280例孕妇血清,TOX-IgM抗体阳性659例,阳性率12.48%。其中妊娠早期阳性率12.24%,妊娠中期阳性率12.97%,妊娠晚期阳性率12.01%。各期TOX-IgM抗体阳性率差异无统计学意义(χ2早、中=0.46,χ2早、晚=0.36,χ2中、晚=0.73,P均>0.05)。659例TOX-IgM抗体阳性孕妇流产或分娩的胎儿组织或脐血TOX-IgM抗体阳性481例,垂直传播率为72.99%。其中妊娠早期垂直传播率48.99%,妊娠中期垂直传播率82.81%,妊娠晚期垂直传播率84.09%。妊娠早期垂直传播率与妊娠中期、晚期比较差异有统计学意义(χ2=14.50,P<0.05),妊娠中期与晚期垂直传播率差异无统计学意义(χ2=0.13,P>0.05)。结论孕妇妊娠各期弓形虫感染的几率相近,而垂直传播率在妊娠早期显著低于妊娠中期和妊娠晚期,提示在确诊孕妇弓形虫感染后,应及时采取干预措施,降低弓形虫感染的垂直传播。     

Seroprevalence of specific total immunoglobulin (Ig), IgG and IgM antibodies to Toxoplasma gondii in blood donors from Loei Province, Northeast Thailand

A total samples from 345 healthy blood donors from Loei Province, Northeast Thailand were examined for anti-Toxoplasma gondii antibodies by ELISA. The seroprevalence of the anti-Toxoplasma gondii total Ig, IgG and IgM antibodies was 4.9%, 4.1% and 4.3%, respectively. Overall seropositive rate was 33 out of 345 individuals (9.6%). Among the seropositve cases, 5 (15.2%), 2 (6.1%) and 13 (39.4%) of the samples were determined by using each type of anti-T. gondii total Ig, IgG and IgM antibodies, respectively. The seropositive sera was also determined by combining of anti-T. gondii antibodies (anti-T. gondii total Ig with IgG and anti-T. gondii total Ig with IgM antibodies). These results were 10 (30.3%) and 2 (6.1%) cases, respectively. Only one (3%) sample had all types of anti-T. gondii antibodies. In addition, the frequency distribution curves of ELISA optical densities of anti-T. gondii total Ig, IgG and IgM antibodies in blood donor presented "unimodal" curves. The negative results were found in the age group that less than 20 years old and more than 51. The highest seropositive results were found in two age groups (21-30 and 31-40 years old), and males were significantly higher than female (p < 0.05). These results demonstrated that when using anti-T. gondii total Ig, IgG and IgM antibodies for determining the seroprevalence, the sensitivity was twice that with the anti-T. gondii, total Ig antibody alone.     

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。     

弓形虫感染者血清IgM和IgG抗体免疫印迹谱分析

目的探索近期和慢性弓形虫感染的免疫诊断标志物。方法采用无毒灵敏的TMB为底物的酶联免疫印迹技术,分析弓形虫感染者IgM或IgG抗体阳性血清与弓形虫速殖子可溶性抗原的免疫反应谱。结果IgM抗体对p35、p38和p22的识别比例达80%以上,且反应带出现率较其它抗原组分高,而p32、p30与IgG抗体的反应带出现率和识别比例也分别高达40%和80%以上。其中,反应最强烈最具代表性的分别属p35(出现率81.5%,P<0.0001)和p32(出现率57.1%,P<0.0001)。结论p35、p38、p22可能为近期弓形虫感染的诊断标志物,p32和p30则可能是慢性弓形虫感染的诊断标志物。