现场应用粪抗原检测法诊断犬细粒棘球绦虫感染的效果评价

目的 评价粪抗原检测法在包虫病流行病学监测中的实际应用价值。方法 家犬在采集粪便后，进行槟榔碱导泻，应用粪抗原检测法检测粪样中的细粒棘球绦虫抗原，并与槟榔碱导泻法的驱虫结果比较，同时分析犬肠道中寄生的其它蠕虫对粪抗原检测结果的影响。结果 在294只槟榔碱导泻成功的家犬中，45只检出细粒棘球绦虫，粪抗原阳性犬共46只。二者的符合率为97．8％。在249只未检出细粒棘球绦虫的家犬中，粪抗原阳性者1只。结论 粪抗原检测法诊断犬细粒棘球绦虫感染具有较高的敏感性和特异性。可作为包虫病常规监测方法推广应用。     

粪抗原检测法诊断犬细粒棘球绦虫感染的研究Ⅲ.双抗体夹心ELISA法检测粪抗原诊断家犬细粒棘球绦虫感染的现场评价

本文报道用槟榔碱驱虫法对检测粪抗原诊断家犬细粒棘球绦虫感染的应用价值进行现场比较评价的结果.73只受试犬中有16只对槟榔碱无反应.在导泻成功的57只犬中粪抗原阳性者11只阳性率19.2%.其中驱虫阴性的38只犬中粪抗原阳性者仅2只(5.3%).在10只驱出细粒棘球绦虫的家犬中8只粪抗原阳性.5只驱出泡状带绦虫的犬中粪抗原阳性者1只,但反应强度低.粪抗原的反应强度与细粒棘球绦虫的感染强度呈正相关(r=0.94).证明本法可用于家犬细粒棘球绦虫感染的调查.     

粪抗原检测法诊断犬细粒棘球绦虫感染的研究Ⅲ.双抗体夹心ELISA法检测粪抗原诊断家犬细粒棘球绦虫感染的现场评价

本文报道用槟榔碱驱虫法对检测粪抗原诊断家犬细粒棘球绦虫感染的应用价值进行现场比较评价的结果。７３只受试犬中有１６只对槟榔碱无反应。在导泻成功的５７ 只犬中粪抗原阳性者１１只阳性率１９．２％ 。其中驱虫阴性的３８只犬中粪抗原阳性者仅２只（５．３％ ）。在１０只驱出细粒棘球绦虫的家犬中８只粪抗原阳性。５ 只驱出泡状带绦虫的犬中粪抗原阳性者１ 只,但反应强度低。粪抗原的反应强度与细粒棘球绦虫的感染强度呈正相关（ｒ＝ ０．９４）。证明本法可用于家犬细粒棘球绦虫感染的调查。     

吡喹酮长效缓释棒控制家犬细粒棘球绦虫感染的现场研究

目的　为确定吡喹酮驱虫棒在包虫病现场控制家犬细粒棘球绦虫感染的效果。方法　以吡喹酮 1 6～ 2 5g/犬剂量 ,给家犬皮下埋植。植前测定犬粪抗原。植后测定实验组和对照组犬粪抗原并同时做槟榔碱导泻的方法进行预防效果评价。结果　Ⅰ号药棒埋植后 8个月 ,粪抗原阳性率从埋药前的 19 5 %下降至 4 6 %。槟榔碱导泻结果 ,实验组的成虫感染率比对照组低 5倍以上 ,差异显著。Ⅱ号药棒埋植前犬群粪抗原阳性率为 4 1 3% ,埋植 1年后实验组粪抗原阳性率为 0 9% ,对照组为 36 4 %。结论　Ⅰ号药棒可以保护未感染犬不发生感染 ,但不能完全驱除已感染并寄生的成虫。Ⅱ号药棒实际上可以完全控制家犬细粒棘球绦虫的感染。这种药棒可以做为包虫病预防措施的主要手段 ,在流行地区推广使用。     

粪抗原检测法诊断犬细粒棘球绦虫感染的研究：Ⅲ.双抗体？…

本文报道用槟碱驱虫法对检测粪抗原诊断家犬细粒棘球绦虫感染的应用价值进行现场比较评价的结果。７３只受试犬中有１６只对槟 无反应。在导泻成功的５７只犬中粪抗原阳性者１１只阳性率１９．２％，其中驱虫阴性的３８只犬只粪抗原阳性者仅２只。     

甘南藏族自治州终末宿主犬感染棘球绦虫状况调查

目的掌握甘南藏族自治州终末宿主犬细粒棘球绦虫和多房棘球绦虫感染状况,为该地区包虫病的传播动力学研究及开展大规模包虫病防治做好前期工作。方法选择碌曲县和玛曲县当地家犬、牧羊犬,采用15%槟榔碱溶液（21mg/kg体重）导泻后检查成虫,随机捕杀当地无主野犬,解剖十二指肠检查成虫。结果终末宿主犬细粒棘球绦虫感染率为22.97%（17/74）,多房棘球绦虫感染率5.41%（4/74）,合计感染率为28.38%,感染犬的感染度为43.29条/只。未发现两种绦虫混合感染。结论甘南藏族自治州犬细粒棘球绦虫感染率较高,并有多房棘球绦虫感染。     

家犬粪便中的细粒棘球绦虫DNA PCR检测方法的建立

目的 建立检测家犬粪便中细粒棘球绦虫DNA的PCR方法，为开展流行病学监测提供检测手段。方法 犬粪经液氮反复冻溶后提取DNA，PCR扩增编码细粒棘球绦虫12srDNA检测该靶DNA片段（255bp）。结果 10只细粒棘球绦虫感染犬，8只（体内成虫数〉80条）阳性，2只（体内成虫数分别为4条和5条）阴性。4只未感染家犬粪样及其他3种绦虫DNA样本均为阴性。结论 初步建立了灵敏、特异的检测家犬粪便中细粒棘球绦虫的PCR方法。     

棘球绦虫感染犬粪抗原检测研究新进展

摘要：棘球绦虫（Echinococcus spp）感染的犬目前是人畜棘球蚴病主要传染源,因此对感染犬的动态监测、合理驱虫治疗是预防和彻底消灭包虫病流行的关键性措施之一。近年来,为了克服槟榔碱导泻法鉴定虫种和虫卵以及检测血清特异性循环抗原及抗体等方法的不足,各实验室开始应用ELISA双抗夹心法,试图建立棘球绦虫感染犬粪抗原的快速检测系统。本文对犬感染棘球绦虫的特异性粪抗原检测技术的最新进展作一综述。     

PCR诊断犬感染细粒棘球绦虫方法检出日期的确定

目的确定犬细粒棘球绦虫感染PCR检测的窗口期。方法家犬8只,口服喂饲细粒棘球绦虫原头节,感染后40 d,收集犬粪,用PCR方法检测粪便中的目的DNA。结果6只犬获得细粒棘球绦虫重度感染,感染后21 dPCR检出目的DNA片段。结论犬重度感染细粒棘球绦虫后粪便PCR检测的窗口期为20 d。     

犬细粒棘球绦虫感染LAMP检出日期的确定

目的确定犬细粒棘球绦虫感染环介导等温扩增技术(LAMP)最早检出日期。方法家犬8只,口服喂饲细粒棘球绦虫原头节,建立细粒棘球绦虫感染动物模型。感染后40d收集犬粪,用LAMP检测粪便中细粒棘球绦虫目的基因并与传统的PCR比较。结果 8只犬均获得细粒棘球绦虫感染,感染后第18dLAMP法检查犬粪中细粒棘球绦虫目的基因均阳性,较普通PCR法提早2d。结论 LAMP法简便、快捷、灵敏,有望成为早期诊断犬细粒棘球绦虫感染的重要手段。     

粪抗原检测法诊断犬细粒棘球绦虫感染的研究 Ⅰ.用细粒棘球绦虫成虫虫体抗原—抗体系统检测粪抗原方法的建立

用细粒棘球绦虫成虫虫体抗原及其免疫家兔血清ＩｇＧ建立了用于检测家犬粪抗原诊断细粒棘球绦虫成虫感染的双抗体夹心ＥＬＩＳＡ方法。对用离子交换层析、ＳＰＡ亲和层析、抗兔ＩｇＧ抗体亲和层析和细粒棘球绦虫成虫虫体抗原亲和层析等四种方法纯化的抗体进行了比较，以抗原亲和层析法纯化的抗体免疫活性最高，用双抗体夹心ＥＬＩＳＡ检测抗原的终点在标本缓冲液中为２．５ｎｇ／ｍｌ，在健康家犬粪便悬液的上清中为５ｎｇ／ｍｌ。检测１１只人工感染细粒棘球绦虫成虫的家犬粪便标本，粪抗原阳性率达１００％。     

槟榔碱测试对犬驱虫管理体系的监测

1989年4月,在和硕县选8个点进行现场调查,对310条犬进行了槟榔碱测试,以评价该县包虫病防治的近期效果。结果表明,该县犬群的登记管理比例由1988年的95％降至1989年的61.98％。1988、1989两年驱虫强度的各项指标分别为:平均驱虫次数2.85和0.85次;平均驱虫密度为35.63％和28.6％;驱虫效率为0.99和0.64。这种驱虫强度在16个月间使全县犬群Eg感染率由30％降至10.4％,但仍维持在全疆农区较高水平。驱虫1～2次的犬与未驱虫犬对Eg感染无显著性差异,阳性犬所占比例随驱虫次数增加而减小。驱虫频率在50天间隔内,犬阳性率可为零。未驱虫犬的相对感染风险是驱虫犬的2.53倍。犬主的民族、职业和犬的用途在犬的Eg感染中有显著差异。 本研究有力地支持了犬6周驱虫的防治对策,证实强化驱虫完全可以抵消社会因素和犬主行为对犬感染的影响,并发现犬同时感染Eg和大绦虫的机会比单独感染Eg的机会高4.99倍。 对以阳性犬来衡量人、畜感染潜在风险的观点进行了讨论。     

粪抗原检测法诊断犬细粒棘球绦虫感染的研究Ⅱ.粪抗原检测的敏感性、特异性及实验感染犬排出粪抗原的动态观察

用细粒棘球绦虫成虫虫体抗原及其抗体建立的双抗体夹心ＥＬＩＳＡ滴定成虫抗原的结果Ａ４５０值与抗原含量呈指数曲线关系,高度相关（ｒ＝０．９８）。实验感染家犬粪抗原含量在２．５－８．０μｇ／ｍｌ之间,因此本法在检测粪抗原中具有足够的敏感性。家犬泡状带绦虫的感染引起的交叉反应是影响本法特异性的主要问题。选择最适的抗体包被浓度可消除交叉反应。用细粒棘球蚴原头节感染家犬后,粪抗原呈低滴度阳性,至２１天后迅速升高。实验感染泡状带绦虫的家犬观察５０天粪抗原均在临界水平之下。作者推荐本法可用于家犬细粒棘球绦虫成虫感染的诊断。     

四川省藏族牧区家犬棘球绦虫病流行病学调查研究

目的 对四川省甘孜县达通玛藏族牧区家犬感染细粒棘球棘球绦虫和多房棘球棘球绦虫进行流行病学调查和评价感染风险因素.方法 分别对甘孜县达通玛藏族牧区查龙、卡龙、大德和查扎等4乡的犬主进行问卷调查,了解家犬感染棘球绦虫的相关因素.剖检流浪犬,检测棘球绦虫感染率,并用此结果 评价粪抗原-ELISA方法 .用该方法 检测家犬感染棘球绦虫的阳性率,评价犬驱虫效果.用X2检验和方差分析对结果 进行统计. 结果 2000年流浪犬棘球绦虫感染率为60.9%(14/23),其中细粒棘球绦虫感染率为26.1%(6/23),多房棘球绦虫感染率为34.8%(8/23).粪抗原-EUSA特异性为80.0%,敏感性为9213%.家犬粪抗原-EUSA阳性率平均为50%(290/580).从2003年起,经每半年1次吡喹酮犬驱虫(5 mg/kg),2005年同一犬群粪抗原阳性率降为17.0%(99/580).犬感染风险因素调查发现敞放犬粪抗原阳性率[40.4%(65/161)]明显高于半栓养犬[白天拴养夜晚放养的犬32.3%(109,337);夜晚拴养白天放养的犬29.2%(21/72)]及一直栓养的犬[20%(2/10)1(P＜0.01);主人缺乏防治相关知识的犬[38.1%(121/318)]和不进行驱虫的犬[47.7%(92/193)],阳性率明显高于主人具有相关知识[28.6%(75/262)]和驱虫犬120.4%(79/387)](P＜0.05和P＜0.01).粪抗原-ELISA阳性率与犬的年龄、性别和饲养家畜的种类无关.结论 四川省甘孜县达通玛藏族牧区是家犬两种棘球绦虫病流行区.粪抗原-EUSA法可用于检测犬棘球蚴病.犬敞放和不对犬驱虫,以及牧民缺乏相关知识是造成家犬棘球蚴病传播、流行的重要原因.     

细粒棘球蚴原头节虫体和排泄分泌抗原抗体系统检测犬粪抗原的比较研究

用细 粒棘球蚴原头节虫体及排泄分泌抗原－抗体系统进行交叉酶联免疫吸附试验的实验结果显示;成虫虫体抗原和原头节虫体抗原呈现交叉反应,表明这两个发育阶段的虫体抗原具有主要的共同抗原组分;原头节排泄分泌抗原的免疫抗原是原头节的期特异性抗原,与其相应抗体具有较高的亲合力。检测犬粪标本的结果感染细粒棘球绦虫的犬粪抗原是一种广谱抗原。两种抗原－抗体系统均可成功地用于粪抗原的检测。     

Copro-diagnosis of Echinococcus granulosus infection in dogs by amplification of a newly identified repeated DNA sequence

Diagnosis of Echinococcus granulosus infection in dogs by detecting adult worms recovered post mortem or purged from the intestines after treatment with arecoline is not suitable for mass screening. Large-scale diagnosis by detection of copro-antigens is useful but only with relatively high intensity infections, and only by genus. To provide a more sensitive and specific diagnosis, a polymerase chain reaction (PCR) assay was developed, that amplified a target repeated sequence (EgG1 Hae III) newly identified in the genome of the common sheep strain of E. granulosus. This repeated sequence consists of approximately 6,900 copies, arranged in tandem, in groups of 2-6 repeats. The corresponding primers used in the PCR easily detected a single egg with no cross-amplification of DNA from closely related cestodes, including E. multilocularis and Taenia spp. Fecal samples from naturally infected dogs, with 2-10,000 E. granulosus worms at necropsy, were all PCR positive, while E. multilocularis or Taenia spp. positive controls as well as non-endemic controls were all PCR negative. This copro-PCR assay was demonstrated to be 100% specific and also detected all necropsy-positive E. granulosus-infected dogs. It is suggested that this copro-PCR assay has the potential for pre-mortem diagnosis of E. granulosus infection even in areas where E. granulosus and E. multilocularis are co-endemic.     

Canine echinococcosis in Turkana (north-western Kenya): a coproantigen survey in the previous hydatid-control area and an analysis of risk factors

A study of Echinococcus granulosus infection in dogs, with risk-factor analysis, was carried out in the endemic area of northern Turkana district, Kenya, using necropsy on 42 strays and a coproantigen-ELISA survey of 161 owned animals. During the post-mortem examinations, 14 (33%) of the necropsied dogs were found infected with E. granulosus, with a mean burden of 540 worms (range=two to 4080 worms). The 26 necropsied dogs that came from the north-western Lokichoggio division--an area where, from 1983 to 1997, there had been a continuous programme of hydatid control--showed a similar prevalence of infection to the other dogs (34.6%) but a significantly lower mean burden, of 53 worms (range=two to 300). Forty-two (26%) of the animals tested for Echinococcus coproantigen were found positive. Although the dogs from the Lokichoggio division were more likely to be coproantigen-positive (29%) than those from the central Kakuma division (20%) or the north-eastern division (18%), the differences were not statistically significant. In questionnaire-based interviews, the owners of the dogs tested for coproantigens were asked about possible risk factors for canine infection with E. granulosus. Women were found to have twice the level of contact with dogs as men. The results of a univariate analysis of the dog-owners' responses revealed six factors that appeared to be significantly associated with a coproantigen-positive dog: non-restraint of the dog (P<0.001); dog fed on raw offal (P<0.001); the improper disposal of slaughter offal (P<0.001); the dog-owner's lack of knowledge about the transmission of echinococcosis (P=0.001); the dog not receiving anthelmintic treatment (P=0.003); and dog age < or =5 years (P=0.01). The results of a multivariate analysis confirmed that lack of dog restraint, access to raw offal, and young age of the dog (< or =5 years) each significantly increased the risk of coproantigen positivity (P, 0.005). Dogs that scavenged from cooking pots, were used to clean babies, had access to the inside of houses, and/or slept indoors appeared, however, to be at no increased risk of coproantigen positivity. The present results are discussed in relation both to older information on the epidemiology and role of human behaviour in the transmission of E. granulosus in Turkana, and the effects of the hydatid-control programme that ran continuously in the north-western division of Turkana between 1983 and 1997.