贵州省7个地区主要山羊流产疫病的流行病学调查

为了解2011—2012年间贵州省山羊5种流产疫病的流行情况,采用虎红平板凝集试验(RBPT)、间接血凝试验(IHA)和酶联免疫吸附试验(ELISA)3种血清学方法对贵州省7个地(州、市)74个养殖场的514份山羊血清进行5种流产疫病抗体的血清学监测,并通过PCR方法对10个规模化养殖场流产死亡母羊子宫、胎儿、肺和肺门淋巴结组织进行羊流产亲衣原体病和山羊传染性胸膜炎的病原核酸进行检测。结果显示:布氏杆菌病、羊流产亲衣原体病、弓形虫病、蓝舌病、山羊传染性胸膜炎的丝状支原体山羊亚种和绵羊肺炎支原体血清抗体阳性率分别为0、1.26%、6.72%、26.95%、2.35%和6.79%;羊流产亲衣原体病、山羊传染性胸膜肺炎的丝状支原体山羊亚种和绵羊肺炎支原体病原核酸检测阳性率分别为0、0、10%。结果表明:目前除布氏杆菌病以外,其余4种山羊流产疫病均在贵州存在不同程度的流行。     

贵州省山羊布鲁氏杆菌病衣原体病蓝舌病的血清学调查研究

为了解贵州省山羊3种流产疫病流行情况,分别从贵州省4个地区共采集194份山羊血清,采用虎红平板凝集试验法(RBT)对194份血清进行布鲁氏杆菌病抗体检测,平均阳性率为0%;采用间接血凝试验法(IHA)对194份山羊血清进行衣原体病抗体检测,平均阳性率为5.7%;采用酶联免疫吸附试验法(ELlSA)对178份山羊血清进行...     

青海省英得尔种羊场种羊流产血清学调查

通过间接血凝试验(IHA)和虎红平板凝集试验(RBPT),对来自青海省英得尔种羊场的353份羊血清进行布鲁氏菌、弓形虫和衣原体三种主要疫病的血清抗体检测。结果表明,18.9%(33/174)的山羊血清和3.9%(7/179)的绵羊血清其衣原体抗体血清凝集效价≥1:16(++),被判为阳性;抗布鲁氏菌和弓形虫的血清抗体均未检测到。     

贵州省猪弓形虫病的血清学调查

为全面了解贵州省猪弓形虫感染情况,对采自9个市（州、地）264个养猪场（户）的2 906份血清用酶联免疫吸附试验（ELISA）检测猪弓形虫抗体,总体阳性率为65.83%,变化范围为27.88%~85.42%。采集65份猪血清做ELISA和间接血凝试验（indirect heamagglutination assay,IHA）比较测定,两种方法总体符合率为58.46%。IHA方法补充检测177份猪血清,弓形虫抗体阳性率为27.68%。血清学调查结果与国内部分省（市）报道相符。     

青海省三角城种羊场部分流产母羊的血清学检测

应用布鲁氏菌病RBPT、衣原体和弓形虫IHA,对来自青海省三角城种羊场发生流产的23只生产母羊血清进行特异性血清抗体的检测,结果检出衣原体阳性血清3份,阳性率为13.04%:未检出布氏杆菌和弓形虫阳性血清。     

青海省门源县山羊衣原体病血清学调查

应用衣原体间接血凝试验(IHA),对青海省门源县山羊进行了衣原体血清抗体检测,共检测山羊血清303份,检出阳性48份,阳性率为15.84%。     

单克隆抗体在动物衣原体病血清诊断上的应用

采用淋巴细胞杂交瘤技术，研制出的抗衣原体单克隆抗体，应用酶联免疫吸附试验（ELISA）方法，对本地区所采得的231份猪血清，128份羊血清，90份牛血清进行检测，结果：猪血清阳性检出率为30．7％，羊为25．8％，牛为32．2％；选猪、羊、牛血清各50份，用已建立的检测衣原体抗体ELISA方法与的间接血凝试验（IHA）进行比较，猪血清ELISA阳性检出率为30％，IHA为26％，ELISA比IHA试验高出4个百分点。牛血清ELISA阳性检出率为32％，IHA为24％，ELISA比IHA试验高出8个百分点。羊血清ELISA阳性检出率为26％，IHA为22％，ELISA比IHA高出4个百分点。     

青海省湟中县某奶牛场奶牛流产病因的血清学调查和分析

用试管凝集法(SAT)、间接血凝抑制试验(IHA)和酶联免疫吸附试验(ELISA)三种方法,对来自青海省湟中县田家寨镇某奶牛场发生过流产群的95份奶牛血清,进行布氏杆菌病、衣原体病和弓形虫病的血清学调查。结果:检出布氏杆菌病阳性血清1份,衣原体阳性血清15份,弓形虫阳性血清2份,血清阳性率分别为1.05%、15.79%和2.11%。初步说明引起该牛场流产的病因为布氏杆菌病、衣原体病及弓形虫混合感染。     

羊流产嗜性衣原体重组ompA蛋白的原核表达及间接ELISA抗体检测方法的建立

原核表达羊流产嗜性衣原体(Chlamydophila abortus,C.abortus)ompA蛋白,包被ompA蛋白抗原,筛选反应条件,成功建立了血清抗体ELISA检测方法。PCR扩增ompA蛋白基因并进行原核表达,经纯化、复性和Western blot鉴定后作为包被抗原,通过反应条件筛选、灵敏性、特异性、重复性检验和临床样品检测,创建了羊流产嗜性衣原体血清抗体间接ELISA检测方法。结果表明,建立pET-28α-ompA阳性质粒并表达约为40 000的ompA蛋白,Western blot显示纯化复性的ompA蛋白具备抗原性与特异性。反应条件筛选结果为ompA蛋白包被360 ng;血清及二抗分别稀释1∶50,1∶5 000并37℃作用1 h;TMB显色10 min。在优化条件下,D_(450)≥0.327为阳性,反之为阴性;特异性、敏感性和重复性结果显示,对布氏杆菌、羊痘、羊口疮、小反刍兽疫和产气荚膜梭菌阳性血清的检测结果均为阴性且50份流产性衣原体阳性血清检出率为98%,批内和批间D_(450)值变异系数分别为1.56%～2.56%和2.35%～3.25%。应用建立的方法对临床175份血清检测,阳性率为52%,商品化衣原体间接血凝检测阳性率为49.1%,阳性符合率为96.5%。本试验建立的间接ELISA检测方法可用于羊流产性衣原体血清抗体水平检测。     

羊布氏杆菌病 衣原体病对比试验诊断

采集119份流产母羊和随机抽检81份母羊血清,作布氏试管凝集和衣原体间接血凝试验诊断。在流产母羊血清中,两试验的阳性率分别为59.66％(71/119),23.53％(28/119);在布病SAT的阳性羊内,混感衣原体病的阳性率为23.94％;在衣原体IHA的阳性羊内,混感布病的阳性率为60.71％。在随机抽检母羊血清中,两试验的阳性率分别为2.47％(2/81),18.52％(15/81);在布病SAT的阳性羊内,混感衣原体病的阳性率为50％;在衣原体IHA的阳性羊内,混感布病的阳性率为6.67％。试验证明,在流产严重羊群中,布病为第一致病因素;当布病接近控制时,衣原体又是第一致病因素。     

集约化养猪场嗜流产衣原体病流行状况的初步调查

本试验对北京郊区5个规模化猪场断奶仔猪、育成猪、育肥猪和怀孕母猪采集血清共507份,同时采集密切接触饲养人员、仔猪、育肥猪的喉头拭子、母猪阴道拭子及公猪的精液共183份。分别使用间接血凝诊断试剂盒、法国ELISA试剂盒检测抗体;利用直接荧光染色法检测抗原,包括166份猪喉头拭子、阴道拭子和精液以及17份饲养人员喉头拭子。结果发现间接血凝诊断试剂盒、ELISA试剂盒抗体阳性率分别为4.14%和2.17%;抗原平均阳性率为14.8%,其中精液阳性率达到37.5%,母猪阴道拭子阳性率为27.5%,饲养人员阳性率为23.5%。本试验证实北京郊区猪嗜流产衣原体在种公猪、母猪群和饲养员呈高流行趋势。同时研究结果显示,5个被调查的规模化猪场嗜流产衣原体抗体阳性率显著低于有报道的其他省市,这与间接血凝诊断试剂盒检测结果高于ELISA试剂有关。针对发现的问题,必须采取有效措施控制种公猪精液污染衣原体现象,阻断向母猪和饲养人员传播,以降低人兽共患病发生的风险。     

Field evaluation of a new commercially available ELISA based on a recombinant antigen for diagnosing Chlamydophila abortus (Chlamydia psittaci serotype 1) infection

A new commercially available ELISA (ELISAr-Chlamydia) for detecting antibodies against Chlamydophila abortus has been evaluated using sheep field serum samples. The ELISA is based on a recombinant antigen which expresses part of a protein from the 80-90kDa family that is specific to C. abortus. Sera (105) from six flocks with confirmed ovine chlamydial abortion (OEA) outbreaks were used in this study, as well as sera (258) from 18 flocks which had suffered no OEA in the last lambing. The ELISAr-Chlamydia was compared with the complement fixation test (CFT) and with an ELISA using purified C. abortus elementary bodies (ELISA-EB), employing as reference technique a comparative microimmunofluorescence test that differentiates C. abortus infection from Chlamydophila pecorum infection. The results showed that the sensitivity of ELISAr-Chlamydia was 90.9% with a specificity of 85.9%, the sensitivity of CFT was 71.0% with a specificity of 83.6%, while the sensitivity of ELISA-EB was 95.2% and the specificity was 54.2%. Furthermore, ELISAr-Chlamydia was the test with fewer false positives resulting from positive reactivity to C. pecorum, although 15% of the sera positive for C. pecorum but negative for C. abortus antibodies reacted positively. This study demonstrated with field material that ELISAr-Chlamydia provides the most balanced results between sensitivity and specificity, especially in flocks with no clinical OEA but reactivity to C. abortus.     

ELISA和IHA检测弓形虫抗体的比较试验研究

目的:分析ELISA方法和IHA方法检测弓形虫病的可靠性。方法:本文应用酶联免疫吸附试验(ELISA)和间接血凝试验(IHA)两种方法,平行检测来自龙岩市某个猪场的32份猪血清中的抗弓形虫特异性抗体。结果:ELISA和IHA对猪弓形虫抗体阳性检出率分别为62.5%(20/32)、53.13%(17/32),这两种方法的阳性检出符合率较高,为71.88%(23/32)。其中,ELISA方法的敏感性(93.33%)、检测效率(81.25%)以及Youden指数(63.92%)都在IHA方法之上,但ELISA方法的特异性(12/17,70.59%)略低于I-HA方法(13/17,76.47%)。结论:ELISA和IHA两种方法均可用于猪弓形虫病的诊断和血清学调查,但ELISA方法更适用于猪弓形虫病的血清学调查。     

应用斑点酶联免疫吸附试验检测水牛伊氏锥虫抗体

应用斑点酶联免疫吸附试验(Dot—ELISA)检测水牛伊氏锥虫IgG抗体,并与间接血凝试验(IHA)进行了比较。检测160份伊氏锥虫疫区水牛血清160份,Dot—ELISA和IHA的阳性率分别为55.63％和53.75％;其中42份虫检阳性血清的阳性率分别为100％和92.86％,两种试验的一致率为95.63％。两种试验的滴度呈显著正相关(r=0.8842)。10份非疫区健康水牛血清两种试验均为阴性。     

Toxoplasma gondii and Chlamydophila abortus in caprine abortions in Tobago: a sero-epidemiological study

A sero-epidemiological study was conducted on a goat farm that experienced an abortion epidemic in the 2005 breeding season in Tobago. Serum samples of goats (aborting and non-aborting) and cats were collected, in addition to the use of stored sera from the farm sampled in 2003 and 2004. Farm records on the reproductive and mortality rates for year 2003, 2004 and 2005 were also reviewed. The sera were screened for Toxoplasma gondii antibodies using the latex agglutination test (LAT), Chlamydophila abortus with an enzyme-linked immunosorbent assay (ELISA) and Brucella abortus using the buffered plate agglutination test (BPAT). Farm records revealed that for the period 2003-2005, the average kid per doe rate decreased from 2.1 to 1.5, the mortality rate increased from 6.3% in 2002 to 19.4% in 2004 and the fertility rate decreased from 98-99% (2002-2004) to 89% (2005). There was a dramatic increase in the abortion rate from <1% (2002, 2003 and 2004) to 29.2% (2005). Of a total of 161 sera tested comprising 12 from 2003, 89 from 2004 and 70 from 2005, 0 (0.0%), 21 (23.6%) and 45 (64.3%) were positive for T. gondii agglutinins (i.e. titres > or =1 : 64) and the differences were statistically significant (P < 0.05; chi(2)). Of all serum samples tested, only 1 (1.1%) of 89 from 2004 was positive for C. abortus while all the sera tested were negative for B. abortus. Amongst the 24 does which aborted in 2005 and were available for testing in mid-2005, 15 (62.5%) had reciprocal titres of > or =1 : 2048, three (12.5%) each had titres of 1 : 1024, 1 : 256 and < or =1 : 16 i.e. negative. The seroprevalence and titres of does that aborted, 20 (87.0%) of 23, all with titres > or =1 : 256 suggesting current infection, were statistically significantly (P < 0.05; chi(2)) higher than was detected amongst does that delivered normal kids, 25 (53.25) of 47 with 22 (48.8%) having titres of > or =1 : 256. One (50.0%) of two cats caught and tested was seropositive with a reciprocal titre of 128. This is considered the first documentation of T. gondii agglutinins in caprine abortion as well the detection of C. abortus antibodies from livestock in Trinidad. It is concluded that of the three zoonotic abortifacient pathogens tested for, T. gondii appeared to have played some aetiological role in the abortion epidemic investigated.     

猪弓形虫血清学检测方法比较

采用弓形虫间接血凝试验（IHA）和弓形虫IgG抗体ELISA两种方法检测来自宁夏区内16个规模场61个散养户的510份猪血清。结果显示,两种方法存在关联性,前者阳性率为19.22%,后者阳性率为22.94%。表明宁夏区内不同类别的养猪场户存在弓形虫病的感染,此外,ELISA方法检测弓形虫抗体检出率高于间接血凝试验。     

Role of Chlamydophila abortus in ovine and caprine abortion in Sardinia, Italy

Between 1999-2003, 14321 sera and 646 abortion samples (498 foetuses and 148 placentae) were analysed from 807 sheep and goat farms distributed all over the island of Sardinia. After notification of abortion in a flock, sera collected at random from adult animals were examined to detect antibodies specific to Chlamydophila (C.) abortus by ELISA, whereas foetuses and placenta were analysed by PCR assay. Specific IgG antibodies were detected in 611 (4.8%) sheep and 106 (5.8%) goats. From a total of 2050 ovine and 151 caprine fetal samples including muscle, liver, abomasum, spleen, brain and placenta, 29 (1.4%) ovine and 1 (0.6%) caprine samples were C. abortus PCR-positive. Placenta was the tissue with the highest detection rate. These results indicate that the seroprevalence of C. abortus infection in sheep and goats is very low in Sardinia, and PCR results demonstrate that C. abortus has no significant role in abortion, especially in goats.     

猪衣原体病的诊断

对湖北省2个猪场的母猪发生流产和返情的疾病进行诊断,采集母猪血样和组织样,对衣原体病、猪瘟等采用IHA、ELISA、PCR等方法进行实验室检验,衣原体IHA抗体、ELISA抗体分别为84.6%(22/26)、88.5%(23/26),PCR检测阳性率为69.2%(18/26)。根据实验室检验结果,结合临床症状分析,诊断为猪衣原体感染,经采取综合防控措施,疫情得到了有效控制。     

贵州省山羊流产的病原学鉴定

为了解贵州省山羊流产与山羊痘的相关性,采用琼脂扩散试验和PCR法对本省10个市(县)流产羊群的血清和病料样本进行山羊痘抗原抗体及病原核酸检测,同时血清进行布氏杆菌抗体检测,流产胎儿病料进行羊流产亲衣原体病原核酸检测。结果发现山羊痘羊群流产率达37.1%(4329/11660),山羊痘血清抗体阳性率为38.2%(34/89),抗原阳性率为72.7%(32/44),流产胎儿山羊痘病毒核酸检出率为83.3%(10/12),发病羊群未检出布氏杆菌和羊流产亲衣原体感染。结果表明,山羊流产与山羊痘感染有一定关系,提示在山羊养殖中应加强饲养管理,防止山羊痘感染引起孕羊流产。