检测日本血吸虫感染者血清中特异性IgG4的诊断价值

为评价日本血吸虫感染者血清中特异性IgG4的诊断和疗效价值，本研究以SEA为抗原，胶体金．抗人IgG4单抗结合物为检测标记物，以金标免疫渗滤法（DIGFA）检测急性和慢性血吸虫病患者治疗前后血清中特异性IgG4抗体。结果显示，急性和慢性血吸虫病病人血清中IgG4阳性率分别为90．9％（30／33）和98．0％（98／100）；检测非疫区健康者血清及其他寄生虫（包括肺吸虫、华支睾吸虫、囊虫等）感染者血清共235人份，未有阳性出现（特异性为100％）；检测急性血吸虫病病人治疗后6个月和12个月血清，IgG4抗体的阴转率分别为52．0％（13／25）和87．5％（21／24），均明显高于IgG阴转率；检测血吸虫病病人治后6个月血清，慢性病人与急性病人IgG4抗体阴转率无差异。结果表明DIGFA法检测病人血清特异性IgG4诊断血吸虫病敏感性高，与IgG相比有更高的特异性，具有一定的疗效考核价值。     

滴金免疫渗滤法检测广州管圆线虫抗体的实验研究

目的建立一种检测广州管圆线虫抗体的简便、快速、敏感、特异的新方法。方法以广州管圆线虫成虫抗原为包被抗原，金标记SPA为显色剂，建立检测广州管圆线虫抗体的滴金免疫渗滤法（DIGFA）；并以ELISA平行检测比较。结果用DIGFA检测广州管圆线虫实验感染鼠血清50份，其阳性检出率为96．0％，50份正常鼠血清的阴性符合率达100％。DIGFA分别检测蛲虫阳性鼠血清13份。绦虫阳性鼠血清11份和粪类圆线虫阳性鼠血清18份，前二者均为阴性，后者交叉反应率为5．6％；DIGFA与ELISA平行检测30份广州管圆线虫阳性鼠血清，两法符合率达96，7％。结论DIGFA与ELISA有相似的敏感性和特异性。且具有简便、快速、不需特殊仪器设备等优点。是一种检测广州管圆线虫抗体的新方法。     

斑点金免疫渗滤试验同步检测抗梅毒螺旋体IgM和IgG抗体的研究

目的探求一种可用于梅毒诊断及疗效考核的简便、快速、可靠的方法。方法建立检测抗梅毒螺旋体抗体的斑点金免疫渗滤试验（DIGFA），平行检测梅毒患者血清中IgM和IgG抗体，对患者在治疗前和治疗后血清中特异抗体水平的动态变化进行研究；以梅毒螺旋体血凝试验（TPHA）法为金标准，对临床上疑为梅毒患者的360份血清进行TPHA法和DIGFA法检测对比。结果用DIGFA法对已确诊为Ⅰ期、Ⅱ期和Ⅲ期梅毒患者治疗前血清中特异IgM抗体的检出率分别为82．61％（38／46）、100．00％（24／24）和77．78％（t4／t8）；IgG抗体的检出率分别为60．87％（28／46）、91．67％（22／24）和88．89％（16／18）；在Ⅰ期梅毒患者血清中特异IgM抗体的阳性率高于I{$抗体（P〈0．05），在Ⅱ期和Ⅲ期梅毒患者血清中，两类抗体的阳性率均较高，且差异无统计学意义（P均〉0．05）。58例两类抗体均阳性的梅毒患者经有效药物治疗后3个月、6个月、1年、1．5年、2年连续采血，经DIGFA检测连续观察的结果表明，IgM抗体出现较早，阴转较为明显和迅速，治疗后3个月的阴转率就达31．03％（18／58），1年的阴转率可达93．10％（54／58）；而IgG抗体出现较晚，阴转缓慢，治疗后1年的阴转率仅为34．48％（20／54）。结论DIGFA是检测抗梅毒螺旋体抗体的有效方法，早期梅毒的诊断和疗效考核可检测IgM抗体，而IgG抗体的检测则主要用于回忆性诊断，不适用于判断是否治愈或再感染。     

斑点ELISA与环幼沉淀试验诊断旋毛虫病的研究

目的比较斑点ELISA法（Dot—ELISA）和玻片环幼沉淀试验（CPT）检测感染旋毛虫大鼠血清抗体的敏感性和特异性。方法采用Dot-ELISA和CPT两种免疫学诊断技术检测实验感染旋毛虫大鼠血清特异性抗体。结果Dot．ELISA和CPT法检测阳性率分别为97．5％和95．O％,差异无统计学意义（x2=0．3463,P〉0．05）。同时用此两种血清学方法检测正常大鼠、斯氏狸殖吸虫感染大鼠、日本血吸虫感染兔及蛔虫病人血清,除1例蛔虫病人CPT阳性外,其他均为阴性。从第2周开始旋毛虫感染大鼠Dot—ELISA和CPT均能测出抗体,第5周达高峰。结论Dot．ELISA和CPT对旋毛虫特异性IgG抗体的检测均有较好的敏感性和特异性。     

日本血吸虫未成熟虫卵可溶性抗原诊断血吸虫病的初步探讨

用日本血吸虫未成熟虫卵可溶性抗原(SIEA)作探针，采用ELISA和ELIB检测血吸虫病人血清抗体，对急慢性血吸虫病人的控出率分别为100％和98．2％，未出现假阳性及交叉阳性反应；治疗后3、6和12个月血清抗体的阴转率分别为32．3％、50．9％和64％。其中抗82，73，67，52，42，38，32，31，28，26．21，18kDa分子的血清抗体消失较快。结果表明，SIEA在血吸虫病的诊断中具有较高的敏感性、特异性和潜在的疗效考核价值。     

清新县流动人口血吸虫病监测效果分析

目的通过对清新县流动人口监测结果的分析,探讨优化输入性血吸虫病人的防控措施。方法对清新县流动人口血吸虫病的监测结果进行统计分析。结果2005年清新县对流动人口67045人进行调查．显示有66％的流动人口居住在该县的城乡结合部或农村地区,来自疫区的人群约占流动人口的12％。2001—2008年．采用常规主动监测1052人、选择性主动监测1456人,血吸虫血清抗体阳性率分别为2．57％（27／1052）和4．81％（70／1456）,两者差异有统计学意义（X2=8．25,P〈0．01）;发现输入性临床诊断慢性病人分别为0例和13例（0．89％）,病原确诊病人分别为0例和2例（0．14％）。显示选择性主动监测对查获输入性病例的效果明显优于常规主动监测。结论来自疫区的流动人口中存在一定数量的血吸虫病患者,为做好输入性传染源的防控工作,现阶段要重点开展选择性主动监测,同时加强被动监测,并通过选择灵敏度较高的检查方法以提高监测效果。     

抗结核LAM、16KD、38KD抗体检测对妊娠结核感染诊断价值评估

目的 探讨抗结核LAM、16KD、38KD抗体检测对妊娠期结核感染诊断价值。方法 采用结核分枝杆菌IgG抗体检测试剂盒（蛋白芯片）检测妊娠结核感染及非结核感染患者的抗LAM、16KD及38KD抗体。结果 21例妊娠结核组LAM、16KD、38KD抗体阳性率（敏感性）分别为81％、76．2％、76．2％，而非结核组（26例）阳性率只有3．9％、7.7％、3．9％，特异性在92％以上。结核组两种和三种抗体阳性率（敏感性）分别为57．2％、52．4％，而非结核组阳性率均为“0”，特异性100％，两组比较差异性有显著性。结论 抗结核LAM、16KD、38KD抗体在妊娠结核病中有较高的敏感性，联合检测抗LAM、16KD、38KD抗体特异性高，方法快速简单，对妊娠结核病诊断有较高的实用价值     

关键时刻怎能感冒—海王银得菲策划推广纪实

目的：探讨菌阳肺结核患者化疗前后其血清结核抗体与痰菌转归及预后的关系。方法：化疗前及化疗期，每月检测血清特异性结核抗体PPD－IgG(ELISA法），同时做细菌学检查（8涂片＋培养）。停药后继续追踪两年。结果：74例中，结核抗体（Anti TB)呈持续强阳性反应者2例（复治），化疗无效。余72例，结核抗体与结核菌之间显示出五种关系。（1）抗体与痰菌同时转阴，占5．56％（4／72）。（2）痰菌转阴，抗体仍阳性，占44．44％（32／72）；（3）化疗后抗体才转阳，占16．67％（12／72）；（4）抗体持续阴性，占26．39（19／72）；（5）抗体先于痰菌转阴，占5．56％（4／72）。痰菌转阴时间：第一、五组最长：初治者第四组时间最短，平均1．46个月；复治比初治转阴时间长；复发病例仅见于二、三组。结论：抗体早转阴者，痰结核菌转阴时间晚；（2）抗体比痰菌晚阴者，有结核复发可能；（3）抗体阴性：初治者，抗结核疗效好，复治者则反之；（4）抗体持续强阳性者，痰菌难以转阴。     

用重组SAG1抗原检测弓形虫抗体

目的探讨重组SAG1抗原对弓形虫IgG和IgM抗体的检测效果。方法用rSAG1作抗原建立免疫印迹方法（rSAG1-WB）,与玻片虫体过氧化物酶免疫染色试验（TSHE）平行检测不同来源血清。结果15例病原学检查阳性小鼠血清和5例免疫兔血清的IgG抗体均为阳性,30例正常小鼠血清和10例正常兔血清均未出现阳性反应。rSAG1-WB检测可疑弓形虫病患者血清阳性率为60．3％（38／63）,献血员血清阳性率为6％（3／50）,与TSHE检测结果（65．1％和4％）均无统计学差异（P〉0．05）。1例IgM强阳性血清和13例IgM弱阳性血清在Western—blot检测中分别出现相应的强阳性与弱阳性反应,50例献血员血清均未出现IgM阳性反应,结果与TSHE一致。结论rSAG1-WB检测弓形虫IgG和IgM抗体均具有高度的敏感性和特异性．与TSHE的符合率高。     

EB病毒两种抗体联合检测在鼻咽癌诊断中的应用

目的探讨EB病毒（epstein—barr virus,EBV）抗体在鼻咽癌早期诊断中的应用价值。方法采用酶联免疫吸附试验（ELISA）测定53例鼻咽癌患者、71例鼻部疾病患者和40例正常体检人群血清EBV相关抗体（EBV VCA—IgA和EA—IgA）。结果3组比较,鼻咽癌组血清VCA—IgA的阳性检出率为79．2％,EA-IgA阳性率为50．9％,与其余两组比较差异有统计学意义（P〈0．01）;鼻咽癌组和鼻部疾病组内VCA-IgA和EA-IgA的阳性率比较差异有统计学意义（P〈0．01）;检测鼻咽癌组两项指标的吸光度均值较另两组高;VCA-IgA单独检测及两项联合检测均具有较高的敏感性（79．2％）,EA-IgA单独检测及两项联合检测均具有较强的特异性（93．8％）。结论联合检测鼻咽癌病人血清的VCA—IgA和EA—gA可兼具两者的性能优势,敏感性较高,特异性较强。对鼻咽癌的早期诊断具有重要的临床应用价值。     

抗日本血吸虫SEA特异性IgY应用于循环抗原检测的研究

本研究应用抗日本血吸虫可溶性虫卵抗原(SEA)的鸡卵黄免疫球蛋白(IgY)建立敏感、特异的检测循环抗原的双抗体夹心酶联免疫吸附试验( ELISA).用SEA皮下多点注射法免疫海蓝鸡,水稀释法制备IgY抗体,以辣根过氧化物酶标记纯化的IgY抗体(IgY-E)和兔抗IgG抗体(IgG-E)分别作为检测抗体,IgY抗体和兔抗...     

全血快速检测日本血吸虫抗体免疫层析法的研制和初步评价

本研究采用抗人IgG单克隆抗体标记红色乳胶微球作为探针,以日本血吸虫虫卵可溶性抗原包被硝酸纤维素膜,运用免疫间接法原理,建立了全血快速检测血吸虫抗体乳胶免疫层析法（WB-DLIA）.以WB-DLIA和ELISA分别检测粪检血吸虫虫卵阳性病人标本,阳性符合率为94.40%（51/54）和96.30%（52/54）;检测正常人标本,阴性符合率分别为96.6%（252/261）和 95.40%（249/261）,两法之间无显著性差异（χ2分别为0.21和0.618,P＞0.05）,Youden指数分别为0.91和0.92;对并殖吸虫感染者全血检测的交叉反应率为43.8%（7/16）,其他寄生虫感染者全血检测的交叉反应率为0（0/29）.研究结果显示WB-DLIA与ELISA具有相似的敏感性和特异性,但前者无需分离血清和任何仪器,结果观察清晰,操作更加简便和快速,且易于保存运输,更加适于临床检测、基层推广和大规模现场筛查.     

Plasma antibody profiles as diagnostic biomarkers for tuberculosis

Two billion people are infected with Mycobacterium tuberculosis, the etiological agent of tuberculosis (TB), worldwide. Ten million to 20 million of the infected individuals develop disease per year. TB is a treatable disease, provided that it is diagnosed in a timely manner. The current TB diagnostic methods are subjective, inefficient, or not cost-effective. Antibody-based blood tests can be used efficiently and cost-effectively for TB diagnosis. A major challenge is that different TB patients generate antibodies against different antigens. Therefore, a multiplex immunoassay approach is needed. We have developed a multiplex panel of 28 M. tuberculosis antigen-coated microbeads. Plasma samples were obtained from over 300 pulmonary TB patients and healthy controls in a country where TB is endemic, Pakistan. Multiplex data were analyzed using computational tools by multivariate statistics, classification algorithms, and cluster analysis. The results of antibody profile-based detection, using 16 selected antigens, closely correlated with those of the sputum-based diagnostic methods (smear microscopy and culture) practiced in countries where TB is endemic. Multiplex microbead immunoassay had a sensitivity and specificity of approximately 90% and 80%, respectively. These antibody profiles could potentially be useful for the diagnosis of nonpulmonary TB, which accounts for approximately 20% of cases of disease. Since an automated, high-throughput version of this multiplex microbead immunoassay could analyze thousands of samples per day, it may be useful for the diagnosis of TB in millions of patients worldwide.     

rSj26-Sj32-Immunogold-IgG-Dot-ELISA用于急性日本血吸虫病的诊断价值

目的探讨rSj26-Sj32-Immunogold-IgG-Dot-ELISA用于急性日本血吸虫病患者的诊断价值。方法利用纯化的rSj26-Sj32融合蛋白和日本血吸虫成虫抗原（SjAWA）建立Immunogold-IgG-Dot-ELISA方法检测急性日本血吸虫病患者血清,并以华支睾吸虫病、卫氏并殖吸虫病、棘球蚴病、乙型肝炎、肺结核病患者和健康人血清作对照,比较检测结果。结果 rSj26-Sj32和SjAWA检测急性日本血吸虫病患者的阳性检出率均为100%,特异性分别为97.67%和95.35%;SjAWA与华支睾吸虫病和卫氏并殖吸虫病血清均存在不同程度的交叉反应。结论纯化的rSj26-Sj32融合蛋白是一种较好的免疫诊断抗原。     

Use of serum antibody and lysozyme levels for diagnosis of leprosy and tuberculosis.

Active tuberculosis (TB) and leprosy are difficult to diagnose early because there are few organisms to detect and the specific immune response does not distinguish between active and inactive disease. We developed an immunoassay for lysozyme to see whether serum lysozyme levels could be used to identify individuals with clinical leprosy or TB. The immunoassay for lysozyme proved superior to standard enzyme assays that were less sensitive and reliable. The lysozyme assay was compared with assays for antibodies to Mycobacterium tuberculosis lipoarabinomannan (LAM) and M. leprae phenolic glycolipid-1. The sera tested were from Ethiopian leprosy (paucibacillary and multibacillary) and TB patients and from healthy Ethiopian and U.S. controls. The lysozyme assay was able to detect more of the individuals with TB (sensitivity, 100% for 19 patients) or leprosy (sensitivity, 86% for 36 patients) than either antibody assay. In particular, lysozyme levels were raised in a higher proportion of the paucibacillary leprosy patients (83% of 17), for whom the antibody assays were less sensitive; the LAM IgG and the phenolic glycolipid-1 IgM levels were raised in only 62 and 44% of 16 patients, respectively. The data suggest that lysozyme measurements may be useful in the diagnosis of mycobacterial infections and other chronic infectious granulomatoses.     

rSj26-Sj32-Immunogold-Dipstick试剂对慢性日本血吸虫病的诊断价值研究

目的研究rSj26-Sj32-Immunogold-Dipstick试剂对慢性日本血吸虫病的诊断价值。方法用rSj26-Sj32融合蛋白和日本血吸虫成虫抗原（SjAWA）Immunogold-Dipstick法检测慢性日本血吸虫病患者血清,同时以华支睾吸虫病、卫氏并殖吸虫病、泡型棘球蚴病、囊性棘球蚴病、乙型肝炎、肺结核患者和健康人血清作为对照,比较两种抗原检测抗体效果的差异。结果该法检测慢性日本血吸虫病的敏感性和特异性分别为92.50%和97.67%,诊断该病的阳性预告值、阴性预告值及诊断效率分别为97.37%、93.33%和95.18%,并且与其他疾病患者血清均无交叉反应。结论 rSj26-Sj32-Immunogold-Dipstick试剂可用于慢性日本血吸虫病的免疫诊断。     

Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing

Conventional phenotypic drug susceptibility testing (DST) methods for Mycobacterium tuberculosis are laborious and very time-consuming. Early detection of drug-resistant tuberculosis (TB) is essential for prevention and control of TB transmission. We have developed a pyrosequencing method for simultaneous detection of mutations associated with resistance to rifampin, isoniazid, ethambutol, amikacin, kanamycin, capreomycin, and ofloxacin. Seven pyrosequencing assays were optimized for following loci: rpoB, katG, embB, rrs, gyrA, and the promoter regions of inhA and eis. The molecular method was evaluated on a panel of 290 clinical isolates of M. tuberculosis. In comparison to phenotypic DST, the pyrosequencing method demonstrated high specificity (100%) and sensitivity (94.6%) for detection of multidrug-resistant M. tuberculosis as well as high specificity (99.3%) and sensitivity (86.9%) for detection of extensively drug-resistant M. tuberculosis. The short turnaround time combined with multilocus sequencing of several isolates in parallel makes pyrosequencing an attractive method for drug resistance screening in M. tuberculosis.     

Development of a rapid,simple dipstick dye immunoassay for schistosomiasis diagnosis

In order to develop a rapid, simple immunodiagnostic assay for schistosomiasis, soluble egg antigen (SEA) of Schistosoma japonicum was conjugated with a blue colloidal dye (D-1) produced in China and used to detect antibodies in the sera of schistosomiasis patients. The antigen-antibody complex was captured by anti-human IgG absorbed onto a nitrocellulose membrane dipstick by means of immunochromatography. The results showed that the sensitivity of the dipstick dye immunoassay (DDIA) was 96.7% in 30 cases of acute schistosomiasis (29/30) and 94.1% (79/84) in 84 cases of chronic schistosomiasis. The specificity of the assay was 96.7% in 60 healthy subjects. Cross-reactions were observed in 10.0% of 20 cases of clonorchiasis and in 70.0% of 20 cases of paragonimiasis. The results were similar to those detected by routine ELISA. In a field evaluation of the DDIA kit, the positive rate of the DDIA was 96.7% in 121 cases of schistosomiasis, compared with 90.1% with the circumoval precipitin test (COPT). The antigen conjugated with dye was stable at room temperature for at least 6 months. The results indicated that the dipstick dye immunoassay provided high sensitivity and good specificity for the detection of schistosomiasis and the assay was rapid, simple and cheap, and did not need any equipment. It was more useful for screening target populations for selective chemotherapy than other immunoassays.     

Improved serodiagnosis of tuberculosis using two assay test.

An antigen capture immunoassay was developed for the detection of mycobacterial antigens in sera from patients with tuberculosis. The assay was evaluated together with an antibody measuring enzyme immunoassay in a clinical trial for serodiagnosis of tuberculosis. Sensitivity of the antibody assay for active pulmonary tuberculosis, including relapsed infections, was 75%, and specificity with other lung diseases was 97%. Sensitivity for extrapulmonary tuberculosis was 84.5% and specificity 84%. Sensitivity of the antigen assay for active tuberculosis was 45% with no false positive reactions. Combination of the results from the two assays increased total sensitivity to 96.5% with a positive predictive value of 0.81 and a negative value of 0.98. The two assay test was relatively simple to perform and offered improved serological diagnosis of tuberculosis over a single antibody test.     

PrimaTB STAT-PAK assay, a novel, rapid lateral-flow test for tuberculosis in nonhuman primates

Tuberculosis (TB) is the most important zoonotic bacterial disease in nonhuman primates (NHP). The current diagnostic method, the intradermal palpebral tuberculin test, has serious shortcomings. We characterized antibody responses in NHP against Mycobacterium tuberculosis to identify immunodominant antigens and develop a rapid serodiagnostic test for TB. A total of 422 NHP were evaluated, including 243 rhesus (Macaca mulatta), 46 cynomolgus (Macaca fascicularis), and 133 African green (Cercopithecus aethiops sabaeus) monkeys at five collaborative centers. Of those, 50 monkeys of the three species were experimentally inoculated with M. tuberculosis. Antibody responses were monitored every 2 to 4 weeks for up to 8 months postinfection by MultiAntigen Print ImmunoAssay with a panel of 12 recombinant antigens. All of the infected monkeys produced antibodies at various levels and with different antigen recognition patterns. ESAT-6 and MPB83 were the most frequently recognized proteins during infection. A combination of selected antigens which detected antibodies in all of the infected monkeys was designed to develop the PrimaTB STAT-PAK assay by lateral-flow technology. Serological evaluation demonstrated high diagnostic sensitivity (90%) and specificity (99%). The highest rate of TB detection was achieved when the skin test was combined with the PrimaTB STAT-PAK kit. This novel immunoassay provides a simple, rapid, and accurate test for TB in NHP.