2012年云南省德钦县鼠疫疫源地调查实验室检测结果与分析

目的 调查了解云南省德钦县是否存在鼠疫自然疫源地,为做好鼠疫预防控制工作提供依据。方法 2012年6-7月,应用现场调查和实验室检测结合的方法在德钦县开展鼠疫调查研究工作。对采集的样本采用间接血凝试验（IHA）、酶联免疫吸附试验（ELISA）、胶体金免疫层析（GICA）3种方法进行鼠疫F1抗原和F1抗体的实验室检测,对旱獭脏器进行鼠疫菌分离培养。结果 捕获的9只旱獭脏器未分离到鼠疫菌,F1抗原检测均阴性;174份犬血清、9份旱獭血清、23份鼠血清、853份鼠心脏浸液进行F1抗体检测,犬血清IHA阳性2份、ELISA阳性16份、GICA阳性3份,其中2份犬血清3种方法检测均阳性,免疫印迹实验证实,能与F1抗原发生特异性显色反应。结论 德钦县检出鼠疫F1抗体阳性犬血清,需进一步调查证实德钦县是否存在鼠疫自然疫源地。     

云南省贡山县鼠疫指示动物血清流行病学调查

目的 调查云南省贡山县鼠疫指示动物血清F1抗体情况,为该地是否存在鼠疫自然疫源地提供依据。 方法 2014年10-11月在贡山县的3个乡镇6个自然村19个村组采集鼠疫指示动物血清,采用间接血凝试验(IHA)、酶联免疫吸附试验(ELISA)、胶体金免疫层析(GICA)3种方法进行鼠疫F1抗体的实验室检测。 结果 共检测指示动物血清293份,其中犬血清288份、猫血清5份。IHA检出阳性血清8份,阳性率为2.73%(8/293),ELISA、GICA结果一致,分别检出阳性血清11份,阳性率为3.75%(11/293)。阳性血清均为犬血清。3种方法进行比较,ELISA阳性率优于IHA(2=7.70, P0.01),与GICA无差异。 结论 贡山县鼠疫指示动物血清学检测阳性,需引起重视并作进一步调查研究。     

新疆维吾尔自治区阿勒泰山地鼠疫疫源地的发现与证实

目的 调查、证实新疆维吾尔自治区（新疆）阿勒泰山地是否存在鼠疫自然疫源地。方法 2021—2022年在阿勒泰山地（青河县、福海县、阿勒泰市）开展鼠疫疫源性调查，采用鼠疫细菌学、血清学、分子生物学对捕获和捡拾的啮齿类动物脏器、血清、心脏浸液、媒介寄生蚤进行实验室检验检测，鉴定分离鼠疫耶尔森菌株的生化表型。结果 鼠疫细菌学检验阿勒泰山地青河县、福海县和阿勒泰市捕获和捡拾的自毙啮齿动物合计115只，其中长尾黄鼠和灰旱獭为优势种，分别占总数的60.87%和20.87%，宿主寄生蚤9组136只，其中长尾黄鼠和灰旱獭寄生蚤的优势种分别为方形黄鼠蚤阿勒泰亚种和谢氏山蚤，分别占宿主寄生蚤的67.74%和100.00%。血清学检测血清样本86份、小型鼠心脏浸液21份、自毙啮齿动物脏器研磨液8份，荧光定量PCR方法检测自毙啮齿动物脏器组织提取核酸DNA 8份。1份长尾黄鼠血清鼠疫F1抗体阳性[间接血凝试验（IHA）滴度为1∶128，酶联免疫吸附试验（ELISA）滴度为1∶256]。1份自毙灰旱獭脏器研磨液鼠疫F1抗原阳性[反相间接血凝试验（RIHA）滴度为1∶64,ELISA滴度为1∶128]。2份自毙长...     

艾滋病抗体血清学检测的两种方法比较:附3年4514份标本的比较

目的探究两种方法检测艾滋病抗体血清的优点,比较两种方法的检测结果相关性及其检测质量。方法选择2014年10月至2016年10月在本中心检测HIV抗体人群血清标本4 514份,分别用酶联免疫法和胶体金法进行HIV抗体初筛,并将其中阳性标本送检进行确证,比较两种方法检测结果的相关性及其检测质量。结果使用酶联免疫法检测结果呈阳性者16例,呈阴性者4 498例,其阳性率为0.35%;使用胶体金法检测结果呈阳性者14例,呈阴性者4 500例,其阳性率为0.31%。其中4份血清样本酶联免疫法检测结果呈阳性而胶体金法检测结果呈阴性,2份血清样本酶联免疫法检测结果呈阴性而胶体金法检测结果呈阳性。经配对χ~2检验,酶联免疫法和胶体金法初筛检测HIV抗体的阳性率χ~2=0.33,P0.05,差异无统计学意义;酶联免疫法和胶体金法两种检测方法初筛检测结果共同阳性者12份,共同阴性者4 496份,总符合率为99.87%。两种检测方法对HIV抗体阳性血清无一例漏检,敏感度均为100%,其中酶联免疫法的特异度为99.91%,胶体金法的特异度为99.96%,效果相近。结论对艾滋病抗体血清进行初筛时可选用酶联免疫法或胶体金法,两者敏感度和特异度相近,总符合率高,可互相替代,可依据受检者实际情况选择,但受检者进行筛查试验后必须进行确证试验,以免误诊。     

鼠疫F1抗体胶体金检测试剂的研制和现场评价

目的 为建立鼠疫诊断和监测所需的快速检测F1抗体水平的方法,研制并评价金标检测试剂.方法 胶体金标记和实验室检测,14个省17个监测点现场监测中检测了动物和人的血清样本4789份.结果 金标检测试剂特异性强,敏感性、稳定性好,现场检测结果与间接血凝方法无差异.结论 鼠疫F1抗体胶体金检测试剂可以用于鼠疫病例诊断和动物监测.     

鼠疫菌感染兔血清中的10种重要抗体的检测分析

目的对鼠疫感染兔血清中的10种重要抗体(Ab)进行检测,寻找除F1外的其他可能用于鼠疫菌感染诊断的新靶标。方法利用一种新的基于上转发光法的十通道免疫层析检测技术(TC-UPT-LF)和另一种经典的间接酶联免疫吸附试验(ELISA),同时检测了54份抗F1-Ag抗体阳性的鼠疫菌感染兔血清样品,对样品中10种靶标抗体的阳性率进行了综合分析。结果在54份鼠疫菌感染兔血清样品中抗体检出阳性率较高的有3个,依次为F1-Ab(100%)、YPO1089-Ab(75%)、YopD-Ab(72%),其余7个靶抗体阳性率都低于50%。结论在鼠疫感染的诊断中,除现有的F1外,YopD极有可能作为新的辅助诊断靶标。     

免疫胶体金渗透技术检测抗卵巢抗体的试验研究

目的建立免疫胶体金渗透技术检测抗卵巢抗体(AOAb)的新方法。方法采用人源卵巢抗原,以胶体金颗粒结合的羊抗人IgG为标记抗体,根据免疫胶体金技术原理,建立免疫胶体金渗透技术法检测人血清抗卵巢抗体。分别检测72份抗卵巢抗体阳性血清和88份健康体检女性血清,并与酶联吸附试验的结果进行对比分析。结果与酶联免疫吸附试验的结果对比,其符合率为88.75%,特异性为90.91%,敏感度为86.11%。交叉试验与重复试验结果显示,免疫胶体金渗透技术测定抗卵巢抗体具有稳定性和特异性。结论免疫胶体金渗透技术可以快速准确地测定血清抗卵巢抗体,便于推广普及。     

免疫比浊法检测梅毒螺旋体抗体的临床应用研究

目的探讨免疫比浊法在检测梅毒螺旋体抗体的临床应用。方法收集2014年1~10月东莞广济医院皮肤科血清标本480份,用免疫比浊法、梅毒螺旋体抗体明胶颗粒凝集试验(TPPA)和酶联免疫吸附试验(ELISA)分别进行检测,阳性结果与TPPA检测不符的标本采用荧光螺旋体抗体吸附试验(FTA-ABS)进行验证;另选择10份TPPA检测滴度为1∶1 280阳性标本,用生理盐水按照1∶40、1∶80、1∶160、1∶320、1∶640、1∶1 280进行稀释,用免疫比浊法和ELISA分别进行检测。结果 TPPA检出阳性标本70份,免疫比浊法检出阳性标本69份,ELISA检出阳性标本73份,4份与TPPA阳性结果不相符的标本经FTA-ABS确认全部为阴性;10份滴度为1∶1 280的阳性标本稀释后用免疫比浊法和ELISA对各个滴度进行检测,其敏感性分别为78.3%和96.7%。结论免疫比浊法、ELISA与TPPA3种方法特异性差异无统计学意义,免疫比浊法可以取代TPPA作为梅毒血清学初筛阳性标本的确认试验在临床上推广应用;敏感性方面免疫比浊法比ELISA低,在检测低浓度标本时差异有统计学意义,免疫比浊法不能取代ELISA作为梅毒螺旋体抗体过筛试验。     

胶体金法和酶联免疫法应用于艾滋病抗体检测的结果分析

目的总结分析胶体金法和酶联免疫法应用于艾滋病抗体检测的结果。方法选择在2018年1月-2019年8月我院接受的2000对婚检对象的血液样本作为研究对象,通过胶体金法和酶联免疫法进行抗体筛查,分析两组检测方法的检查结果。结果在作为样本的2000对婚检对象的血液样本中,经酶联免疫法检测,总共有15例阳性,占总比的0.38%,3985份血液样本检测结果为阴性,占总比的99.62%。经胶体金法检测,其中14例为阳性,占总比的0.35%,3986份血液样本检测结果为阴性。两种检测结果相比较,差异无统计学意义(P>0.05)。结论通过将胶体金法和酶联免疫法应用于艾滋病抗体检测,能检测个体HIV病毒感染,可以对临床治疗方案提供参考数据,两种检测方法具有较高的应用价值。     

2000-2007年浙江省云和县鼠疫监测结果分析

目的评价2000-2007年浙江省云和县鼠疫监测结果。方法在原鼠疫流行地区设立固定和流动监测点,开展室内外鼠密度、鼠体蚤和地面游离蚤的监测;并对监测中捕获的活鼠采集脏器样品进行细菌分离培养,同时采鼠血进行鼠疫F1抗体的检测。结果室内、外平均鼠密度分别为3.50％、3.60％;平均染蚤率为8.55％、总蚤指数为0.130;室内、外主要鼠种分别是褐家鼠和黑线姬鼠;鼠体蚤主要为不等单蚤和缓慢细蚤,本地未检获印鼠客蚤。细菌分离培养3551只活鼠,未分离到鼠疫菌;间接血凝试验法检测鼠血清6511份,结果均为阴性;用放射免疫沉淀试验法和金标法检测鼠血清11份,检出1份放射免疫沉淀试验阳性鼠血清。结论通过8年的系统监测,虽未分离到鼠疫菌,但检出放免阳性鼠血清1份,提示云和县可能存在鼠疫的疫情隐患,要加强对鼠疫的监控工作,以防止鼠疫的发生。     

2003-2012年云南省景洪市鼠疫监测结果分析

目的了解和掌握云南省景洪市鼠疫疫情现状,为制定鼠疫防治措施提供科学依据。方法 2003-2012年间,在全市范围内每年设固定监测点1个,每月开展监测,流动监测点每月1~2个,开展鼠疫宿主及媒介监测。宿主动物监测采用笼(夹)夜法,常规方法梳检鼠体寄生蚤,地面游离蚤采用粘蚤纸法捡集。所获鼠血清进行间接血凝试验(IHA)检测鼠疫F1抗体;鼠脏器、蚤标本分别进行鼠疫细菌学分离培养。结果 2003-2012年共捕获鼠5种5657只,鼠密度在2.30%~6.57%之间,总鼠密度4.01%,其中黄胸鼠占99.82%(5647/5657)。染蚤鼠数为753只,染蚤率4.06%~25.34%之间,总染蚤率13.31%,获蚤1784匹,蚤指数在0.11~0.80之间,鼠体总蚤指数0.32。其中印鼠客蚤934匹,占52.35%,黄胸鼠印鼠客蚤总指数0.17(934/5647)。共布放粘蚤纸45 473张,获蚤3种7526匹,平均地面游离蚤指数为0.17。其中印鼠客蚤地面游离蚤指数在0~0.034之间。对5657只剖检的鼠脏器进行鼠疫细菌学分离培养,检出鼠疫菌6株,检菌率为0.11%;IHA 2896份,阳性16份,阳性率为0.55%。反向血凝试验(RIHA)108份,阳性4份,阳性率为3.7%。鼠体蚤、地面游离蚤标本经鼠疫细菌学检验,均为阴性。结论景洪市动物间鼠疫仍时有发生,应加强监测,定期开展群众性灭鼠灭蚤及健康教育工作,有效控制动物鼠疫发生,防止波及人间。     

两种方法测定乙型肝炎病毒血清标志物的比较研究

目的：探讨GIGA （金标快速检测板法）检测乙肝五项血清标志物的结果可靠性。方法357例待测血清标本同时采用ELISA（酶联免疫吸附法）和GIGA（金标快速检测板法）进行检测,对检测结果进行方法学分析。结果357例血清标本中,286例GIGA法与ELISA法结果相符,有71例结果不完全一致。36例ELISA法检测 HBsAg阳性标本中,GIGA法有1例未检出。结论乙肝五项血清标志物的结果GIGA法比ELISA法准确性稍低,但GIGA法操作简便,快速,易于观察结果,且GIGA法中HBsAg检测结果与ELISA法基本一致,适用于急诊手术前以及献血前的快速筛查。     

Antibodies to granulocytes in patients infected with human immunodeficiency virus.

Individuals who are infected with human immunodeficiency virus (HIV) are known to have a high incidence of autoantibodies. In this study, serum samples from 100 individuals with HIV infection were tested for granulocyte antibodies (red cell antibodies, lymphocytotoxic antibodies, circulating immune complexes, and serum immunoglobulin G levels) by granulocyte agglutination (GA) and granulocyte immunofluorescence (GIF) assays. Granulocyte antibodies were detected in 66% of serum samples by GIF and in 21% of serum samples by GA. None of the positive sera reacted with granulocyte antigens of known specificity. Antibodies that reacted with red cell antigens other than ABO were detected in only three serum samples, but lymphocytotoxic antibodies were detected in 62% of patients. All serum samples were tested by immunoblotting with granulocyte plasma membranes. Only two samples were found to be positive; one sample reacted with a 58 kd protein and one reacted with a 55 kd protein, but neither serum sample immunoprecipitated any protein from granulocytes that were labeled at the cell surface with iodine 125. Since immune complexes that are bound to granulocyte membranes can be detected by GIF, circulating immune complex levels were measured in all 100 samples. Immune complexes were increased in GIF-reactive serum samples compared with GIF-nonreactive serum samples (23.3 +/- 19.5 micrograms Eq/ml [mean +/- SD] vs 9.6 +/- 8.1 micrograms Eq/ml, p less than 0.001) but not in GA-reactive serum samples compared with GA-nonreactive sera (24.4 +/- 21.3 micrograms Eq/ml versus 16.9 +/- 16.0 micrograms Eq/ml, p = 0.10).(ABSTRACT TRUNCATED AT 250 WORDS)     

胶体金免疫层析技术同步检测甲、戊型肝炎病毒IgM抗体的研究

目的 建立胶体金免疫层析(GICA)同步检测血清中的甲、戊型肝炎病毒的IgM抗体的方法。方法 采用胶体金标记鼠抗人IgM单克隆抗体，制成免疫层析测试条。血清中甲、戊型肝炎的特异性抗体与测试条上金标抗体结合后沿着硝酸纤维素膜移动与膜上固相包被的抗原(HAAg、HEAg)、抗体(羊抗鼠IgM)结合形成肉眼可见的红色线条。结果 自制的GICA试条与EIA对比检测血清标本188份，各单项指标IgM—抗HAV、IgM-抗HEV两法总符合率依次为97．9％、98．4％，检测灵敏度可达2ng／ml。结论 用于同步检测血清中的甲、戊型肝炎病毒特异性抗体的GICA试条的制备条件已基本建立；其检测结果特异性强、灵敏度高、操作方法简便快捷且无需特殊仪器设备。     

Evaluation of a colloidal gold immunochromatography assay in the detection of Treponema pallidum specific IgM antibody in syphilis serofast reaction patients: a serologic marker for the relapse and infection of syphilis

Syphilis remains as a worldwide public health problem; hence, it is necessary to develop a new diagnostic approach that is easier and faster than conventional tests. A new testing method to detect Treponema pallidum IgM (TP-IgM), named colloidal gold immunochromatography assay (GICA), is presented in place of fluorescent treponemal antibody absorption (FTA-Abs). TP-IgM was detected using GICA developed on syphilis-specific recombinant proteins TPN17 and TPN47. The FTA-Abs IgM test was set as the gold standard. A GICA TP-IgM test was performed to detect syphilis in 1208 patients who received recommended therapy for syphilis for more than 1 year at the Xiamen Center of Clinical Laboratory in China from June 2005 to May 2009. One hundred blood donors were set up as control. The sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio, and negative likelihood ratio were 98.21%, 99.04%, 93.75%, 99.73%, 102.3, and 0.018, respectively. Detection on 500 interference specimens indicated that the biological false-positive rate of the GICA test was extremely low and was free from other biological and chemical factors. The patients were divided into the following experimental groups based on the results of toluidine red unheated serum test (TRUST) and treponemal pallidum particle agglutination (TPPA): (1) the syphilis serofast reaction (SSR) group consisted of 411 cases with (+) TRUST and (+) TPPA, which exhibited no clinical manifestations of syphilis after 1 year of recommended syphilis treatment; (2) the serum cure group, which was further subdivided into group A, a group that consisted of 251 cases with (-) TRUST and (+) TPPA, and (3) group B, a group that consisted of 546 cases with (-) TRUST and (-) TPPA; and (4) the blood donor control group, which consisted of 100 healthy persons with (-) ELISA-TP and (-) TPPA. We used the FTA-Abs method and the GICA method to detect TP-IgM; the positive rate of TP-IgM in 411 SSR patients was 34.55% and 36.01%, respectively. However, in serum cure group A, the positive rate of TP-IgM was 10.36% and 11.16%, respectively. The χ(2) test revealed that there is a significant difference in the positive rate between these 2 groups (P < 0.01). The TP-IgM positive rate in the same group, as detected by the GICA method and the FTA-Abs method, had no significant difference in statistics. However, as detected by the GICA method and the FTA-Abs method, all the samples in serum cure group B and the control group were negative for TP-IgM. The TP-IgM-positive result demonstrated that active T. pallidum remained in the bodies of SSR patients. In summary, the characteristics of GICA TP-IgM correspond to that of FTA-Abs TP-IgM; this can be used as a serologic marker for the relapse and infection of syphilis in place of the conventional FTA-Abs IgM test.     

定量测定HCG的斑点金免疫渗滤试验的临床评价

目的　评价斑点金免疫渗滤试验 (DGIFA)定量测定血清和尿液人绒毛膜促性腺激素 (HCG)的可靠性。方法　分别用化学发光免疫分析法 (CLIA)和DGIFA定量测定HCG试剂盒 (HCG DOT)及DGIFA定量仪测定 4 0份血清样品的HCG含量 ,分别用胶体金免疫层析定性试验 (GICA)和HCG DOT及DGIFA定量仪测定2 70份尿液样品的HCG水平 ;作回收试验和不精密度试验。结果　CLIA和DGIFA定量测定血清样品HCG的相关性好 ;GICA检测阴性的 16 2份尿液样品用HCG DOT测定有 9份为背景红色所致的假阳性 ,1份为血尿导致的假阳性 ,另 2份为阳性 ;GICA检测阳性的 95份和可疑阳性的 13份尿液样品用HCG DOT测定 ,结果均为阳性 ;HCG DOT定量测定高、低浓度HCG血清样品和HCG尿液样品的变异系数 (CV)分别为 5 .38%、4 .13%和 4 .73%、6 .0 1% ;HCG DOT定量测定血清和尿液的HCG回收率分别为 94 .7%和 89.9%。结论　DGIFA定量测定尿液HCG ,对尿液GICA结果可疑者可在妊娠早期作出明确的判断 ;DGIFA定量检测血清和尿液HCG的精密度良好 ,回收率较高 ,适合门急诊应用。     

Detection by flow cytometry of antibodies against surface and intracellular granulocyte antigens

A flow cytometric technique was used to detect granulocyte antibodies, with attention to the distinction between antibodies directed against surface and intracellularly expressed antigens. Ten serum samples with positive results and 10 with negative results detected by the granulocyte immunofluorescence test (GIFT), together with 10 positive serum samples detected by an indirect immunofluorescence test (IIF) were analyzed against leukocytes from healthy blood donors tested by flow cytometry (FC). Unpermeabilized and permeabilized cells were used to allow identification of surface and intracellular binding, respectively. The results of testing by GIFT corresponded with those by FC, with the exception of the results for four sera: one serum sample was FC negative, GIFT positive, and three samples were FC positive, GIFT negative. The IIF-positive sera were all FC positive, analyzed against permeabilized granulocytes, and one serum was also positive against non-permeabilized granulocytes. Analysis by FC is a readily performed technique, which can be used for the routine detection of antibodies against leukocyte antigens. Screening for granulocyte-specific antibodies can be carried out with pools of granulocytes from three donors. Analysis by FC allows detection of both HLA antibodies and granulocyte-specific antibodies and by using both unpermeabilized and permeabilized cells, antibodies against surface and intracellular antigens, respectively, can be identified.