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猪带绦虫六钩蚴cDNA文库的构建与筛选 总被引：1，自引：0，他引：1
在体外将猪带绦虫虫卵孵化为有活力的六钩蚴。采用商品化试剂盒抽提六钩蚴总 RNA、m RNA,反转录为双链c DNA,再加 Eco R Adapter。在去除小片段后 ,dsc DNA与 λgt11噬菌体 DNA连接 ,经蛋白包装 ,构建了猪带绦虫六钩蚴λgt11c DNA文库。该文库效价为 3.5× 10 9pfu/ m L ,重组 DNA片段大小为 0 .5～ 3.2 kb,平均为 1.6 kb,蓝白斑比 1∶ 9。用抗体探针对文库进行免疫学筛选 ,在消除非特异性反应的基础上筛选约 10 6 重组子 ,共得到 118株强阳性克隆 ;应用 PCR鉴定上述部分阳性克隆 ,均扩增到 0 .5 kb以上的片段。结果显示 ,构建的文库合格 ,含六钩蚴所有抗原基因 ,可用于六钩蚴 c DNA克隆的筛选 ;用免疫学与 PCR联合筛选 c DNA文库 ,可消除假阳性 相似文献
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为了研究猪囊尾蚴乳酸脱氢酶(LDH)的表达,试验选择四川省雅江县呷拉乡猪带绦虫病患者,给其口服槟榔-南瓜子驱虫,收集、制备虫卵悬液(8万个/mL),再给3头20日龄三元杂交乳猪猪灌胃虫卵悬液,每头1 mL,40 d后收集、制备猪囊尾蚴蛋白,进行双向电泳(2-DE)分析,将凝胶蛋白斑点转移至聚偏氟乙烯膜(PVDF膜),用自制的大鼠抗猪带绦虫LDH血清作为一抗、健康大鼠血清作为阴性对照进行蛋白质印迹(Western-blotting)分析。结果表明:双向电泳凝胶共检测到(207±9)个蛋白质斑点,相对分子质量(Mr)为14 400～94 000,等电点(pI)为3.0～10.0;试验组特异性抗原抗体阳性杂交斑点为1个,阴性对照未见阳性杂交斑点;将Western-blotting检测的抗原抗体阳性杂交斑点与原双向电泳凝胶斑点进行比对,找到对应蛋白斑点,经ImageMaster 2D Platinum 5.0软件分析后初步确定该蛋白斑点的pI/Mr为7.03/35 368,与猪带绦虫LDH的pI/Mr理论推导值接近,说明猪囊尾蚴表达LDH。 相似文献
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应用免疫组化ＡＢＣ法研究了催产素和加压素在犬泡状绦虫的定位和分布。在犬泡状绦虫中枢神经系统的脑神经节、２条主要纵神经索和环神经内观察到大量阳性纤维和少量阳性细胞。周围神经系统可见阳性神经纤维在皮层和实质形成神经网。从纵神经索和环神经发出的阳性纤维向生殖器官和皮层的肌肉分布。顶突的小钩之间的肌肉存在阳性反应物，小钩周围有较明显的阳性反应环。生殖器官呈现着色深浅不同的阳性反应，以卵囊—梅氏腺复合体最为明显。结果表明，催产素和加压素的免疫反应物具有相同的分布，它们可能参与虫体的神经内分泌调节。 相似文献
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Risk factors for the prevalence of porcine cysticercosis in Mbulu District, Tanzania 总被引：1，自引：0，他引：1
Ngowi HA Kassuku AA Maeda GE Boa ME Carabin H Willingham AL 《Veterinary parasitology》2004,120(4):275-283
To estimate prevalence of and risk factors for the prevalence of porcine cysticercosis in Mbulu District, Tanzania, 770 live pigs were examined by lingual examination in 21 villages. Structured observations and questionnaire interviews were used to assess pig rearing practices and household use of latrines. Associations between factors were analyzed using a Bayesian hierarchical model to obtain prevalence odds ratio (OR) and 95% Bayesian Credible Intervals (95% BCI). Prevalence was 17.4% (village-specific range 3.2–46.7%). Prevalence of porcine cysticercosis was considerably higher in pigs reared in households lacking latrines than in those reared in households that were using latrines (OR=2.04; 95% BCI=1.25, 3.45). About 96% of the pigs were kept under free-range conditions. This study suggests the need for further studies in order to design and implement effective prevention and control measures for porcine cysticercosis in Mbulu District, Tanzania. 相似文献
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Cysticercosis is due to the establishment of the larval stage of the zoonotic cestode parasite Taenia solium. The infection causes substantial human morbidity and mortality, particularly in several Latin American countries and parts of Africa and Asia, as well as economic losses in pig husban dry due to condemnation of infected pork meat. The life cycle of T. solium includes human beings as definitive hosts and pigs as intermediate hosts. Cysticercosis is acquired by the ingestion of eggs released by human tapeworm carriers, who become infected after ingesting pork meat contaminated with cysticerci. Taenia solium transmission has been associated with poverty, lack of sanitary services and practices of rearing backyard pigs with free access to the areas that villagers use as toilets, as well as cultural behaviour. Nonetheless, due to the recent increase of migration and tourism, industrial countries are also reporting cases of human cysticercosis. There are many epidemiological studies that have been conducted mainly in Latin American countries that have evaluated intervention measures for control of cysticercosis including the development and testing of vaccines. Furthermore, the involvement of international agencies and institutions, such as the World Health Organization, the Food and Agriculture Organization and the International Livestock Research Institute, as well as the commitment of policymakers, scientists and field workers, are key means for the sustainable control and, hopefully, eradication of T. solium infections. 相似文献
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RT-PCR扩增45W-4B和TSOL18基因,PCR截去45W-4B基因的N端信号肽和C端疏水氨基酸序列形成45W-4BX。将45W-4BX和TSOL18 PCR产物分别亚克隆人pGEX-4T-1,用IPTG诱导表达,取产物进行SDS-PAGE和Western blot分析。纯化表达产物制成油佐剂疫苗分别免疫家猪,用25000枚猪带绦虫成熟虫卵进行攻击感染,ELISA检测各组的抗体水平,90d后剖检计算各组的减虫率。结果表明,45W-4BX和TSOL18基因在大肠杆菌中分别以可溶性和包涵体形式获得高效表达,并能被囊虫病人血清所识别。重组蛋白免疫猪15d血清抗体即为阳性,30d左右达到峰值。2种重组抗原的减虫率均在88％以上,与囊虫粗抗原免疫效果相当。这为进一步研制基于45W-4BX和TSOL18的猪囊虫重组基因工程疫苗奠定了基础。 相似文献
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Prevalence and seasonal incidence of larval and adult cestode infections of sheep and goats in eastern Ethiopia 总被引：1，自引：1，他引：0
A study on the prevalence and seasonal incidence of cestode parasite infections of sheep and goats was carried out in eastern Ethiopia for 2 years (May 2003-April 2005). During this period, viscera including liver, lungs, heart, kidneys and the gastro-intestinal tract were collected from 655 sheep and 632 goats slaughtered at four abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga. At the abattoirs the abdominal, thoracic and pelvic cavities as well as the muscle surfaces of all animals were visually examined for the presence of larval (cystic) stages of cestode parasites. The viscera were transported within 24 h to the parasitology laboratory of Haramaya University and were examined for larval and adult cestodes following standard procedures. The most prevalent metacestodes (larval cestodes) were Cysticercus ovis (Taenia ovis), Cysticercus tenuicollis (T. hydatigena) and hydatid cysts (Echinococcus granulosus). In sheep, the overall prevalence was 26% for C. ovis, 79% for C. tenuicollis, and 68% for hydatid cysts. Similarly, for goats, the corresponding prevalence was 22%, 53% and 65%, respectively. The difference between sheep and goats in prevalence of C. tenuicollis was significant. The high prevalence of hydatid cysts in both sheep and goats indicates that cystic echinococcosis/hydatidosis is a public health problem in these regions which requires implementation of control measures, including public health education, strict meat inspection and control of stray dogs. The results of the survey also implies that infections of small ruminants with these metacestodes are responsible for condemnation of substantial quantities of affected organs and muscles and therefore of direct economic importance. Intestinal infections with adult tapeworms of Moniezia expansa, Avitellina centripunctata and Stilesia globipunctata, and bile duct infections with Stilesia hepatica were also common in both sheep and goats. In sheep, the overall prevalence of these tapeworms were 61%, 20%, 24% and 39%, respectively. Similarly, the overall prevalence of these parasites in goats was 53%, 21%, 27% and 36%, respectively. 相似文献
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Risk factors associated with porcine cysticercosis in selected districts of Eastern and Southern provinces of Zambia 总被引：1，自引：0，他引：1
Sikasunge CS Phiri IK Phiri AM Dorny P Siziya S Willingham AL 《Veterinary parasitology》2007,143(1):59-66
To determine the risk factors associated with Taenia solium transmission in humans and pigs in the rural areas of Eastern and Southern provinces of Zambia, a questionnaire was administered in 788 households from 155 villages. Pigs were examined from 800 households. Tongue examination and enzyme-linked immunosorbent assay (Ag-ELISA) for the detection of circulating antigens of T. solium cysticerci were used to measure infection in pigs. A snowballing technique was utilised to select households with pigs. Prevalence of households with pigs infected with T. solium on tongue examination by district ranged from 12.7% to 32.1% with Ag-ELISA having a range of 30.0-51.7%. Of the total number of households visited, 18.8% and 37.6% had at least one pig positive for porcine cysticercosis on tongue examination and Ag-ELISA, respectively. Risk factors associated with T. solium infection were lack of pork inspection at slaughter (96.7%), consumption of pork with cysts (20.1%), selling of pork infected with T. solium cysticerci (18.3%), free-range husbandry system (83.2%) and absence of latrines (58.0). Free-range husbandry system (OR=1.68; 95% CI=1.36-2.07) was a significant risk factor for porcine cysticercosis in the surveyed areas. The result that pigs were mostly kept on free-range and semi-intensive husbandry systems may have permitted them to have access to eating human faeces that could be contaminated with tapeworm eggs. This study has shown that T. solium infection poses a high public health risk in the study areas and urban areas as well. We recommend that a human survey be conducted to verify the human exposure to taeniasis and/or cysticercosis in Zambia. 相似文献