弓形虫P30基因及其功能

弓形虫 P30基因所编码的蛋白为弓形虫主要表面抗原 SAG1 ,其约占速殖子总蛋白量的 5 %,SAG1对虫体侵入宿主细胞及其毒力具重要性 ,并有高度免疫原性和免疫保护性 ,被用于弓形虫疫苗的研制和弓形虫感染的分子诊断     

Pathologic and immunohistochemical findings in an outbreak of systemic toxoplasmosis in a mob of red kangaroos

Toxoplasma gondii is a zoonotic protozoan pathogen that infects many endothermic vertebrates, including humans; the domestic cat and other felids serve as the definitive host. Macropodids are considered highly susceptible to toxoplasmosis. Here, we describe the clinical, pathologic, and immunohistochemical findings of an outbreak of systemic toxoplasmosis in a mob of 11 red kangaroos (Macropus rufus), with high morbidity (73%) and mortality (100%) rates. Affected animals had either severe and rapidly deteriorating clinical conditions or sudden death, which was correlated with widespread necrotizing lesions in multiple organs and intralesional T. gondii organisms identified via MIC3-specific immunohistochemistry and confirmed by REP529-specific rtPCR. Quantification of parasites demonstrated the highest parasite density in pulmonary parenchyma compared with other tissues. Our study highlights the continued importance of this severe condition in Australian marsupials.     

孕妇弓形虫病防治研究进展

弓形虫病是一种世界性分布的人兽共患寄生虫病,在人和动物中广泛传播。孕妇感染弓形虫后多表现为隐性感染,无明显的临床表现,但该虫可通过垂直传播影响胎儿发育,造成早产、流产、畸形和死胎等不良妊娠现象。论文对弓形虫病流行病学、感染因素、免疫学、诊断和治疗等方面阐述弓形虫感染对孕妇的影响,为孕妇弓形虫病的防治提供参考。     

Risk factors associated with sero-positivity to Toxoplasma gondii in captive neotropical felids from Brazil

From September 1995 to February 2001, blood samples were collected from 865 neotropical felids belonging to 8 different species. These animals were housed in 86 institutions located in 78 cities of 20 Brazilian states. Our goal was to identify the risk factors associated with sero-positivity to Toxoplasma gondii in captive neotropical felids from Brazil. All serum samples were tested by the modified agglutination test (MAT), using formalin-fixed whole tachyzoites and mercaptoethanol. For each animal an individual questionnaire was filled with questions about tattoo number, felid species, age, sex, origin, number of animals in the group, introduction of new animals in the group, time in the institution, eating meat previously frozen for a period <7 days in the last 6 months, eating meat of run-over or euthanized animals in the last 6 months, predation of rodents or birds in the last 6 months and presence of domestic cats near the enclosures in the last 6 months. The total sero-prevalence was 55% (95% CI: 52%, 57%). We estimated a prevalence of 46% (95% CI: 40%, 54%) for jaguarundi (Puma yagouaroundi); 58% (95% CI: 53%, 63%) for ocelot (Leopardus pardalis); 50% (95% CI: 45%, 56%) for oncilla (L. tigrinus); 54% (95% CI: 46%, 62%) for margay (L. wiedii); 12% (95% CI: 4%, 31%) for Pampas-cat (L. colocolo); 83% (95% CI: 65%, 93%) for Geoffroy's-cat (L. geoffroyi); 64% (95% CI: 50%, 68%) for jaguar (Panthera onca) and 48% (95% CI: 42%, 54%) for puma (Puma concolor). Multiple logistic regression was used to evaluate the association between the variables in the questionnaire and sero-positivity to T. gondii. We concluded that the independent risk factors for toxoplasmosis were: age >3 years (OR = 4.75 [2.75; 8.2]), eating meat previously frozen for a period <7 days (OR = 2.23 [1.24; 4.01]), and consumption of animals that were run-over or euthanized (OR = 1.64; [1.14; 2.37]).     

弓形虫微线体蛋白MIC3基因的克隆及原核表达

根据编码MIC3的已知基因序列设计并合成一对引物,应用PCR技术从弓形虫RH株的基因组DNA中扩增编码MIC3的全长基因,克隆入pGEX-KG表达载体,转化入E.coli DH5α感受态细胞,经含氨苄青霉素琼脂平板筛选,小量抽提质粒进行酶切、PCR及DNA测序鉴定.然后阳性重组质粒转化入E.coli BL21-CodonPlus,IPTG诱导,表达产物经SDS-PAGE和Western-blot分析鉴定.结果显示,扩增的MIC3基因与GenBank中相应基因序列（AJ132530）的同源性达99.6 %,表达的MIC3融合蛋白表观分子量约为66 ku,且可被兔抗弓形虫免疫血清识别.说明所获得的表达蛋白质具有一定的反应原性,为下一步利用重组蛋白建立弓形虫的诊断方法和研制弓形虫的亚单位疫苗奠定了基础.     

北京地区犬猫弓形虫病血清学调查

比较了酶联免疫吸附试验（ELISA）和乳胶凝集试验对犬猫进行龚地弓形虫（Toxoplasma gondii）抗体检测的结果.在66只犬猫的血清中,ELISA方法检测的阳性率为24.0%,乳胶凝集试验阳性率为21.0%,二者的检测结果差异不显著（P＞0.05）.后采用ELISA方法对北京地区家养的159只犬和128只猫进行了弓形虫IGg抗体的检测,共有21只犬（13.21%）和18只猫（14.06%）检测结果为阳性.1岁以下犬（3.70%）、猫（4.35%）低于1至3岁犬（11.29%）和猫（6.98%）的感染率.3～6岁犬猫的感染率分别是27.27%和16.22%,6岁以上犬（30.0%）、猫（32.0%）的感染率最高.不同性别之间感染无明显差异（P＞0.05）.     

弓形虫两种保种方法的试验

目的寻求一个简便、适宜的弓形虫保种方法。方法采用不同温度的冷冻保存法和细胞培养保存法对弓形虫保存进行了试验研究,为深入研究弓形虫提供保证。结果弓形虫在液氮(-196℃)、-70℃能长期保存,4℃能保存14 d。通过细胞培养能获得大量的弓形虫滋养体,对培养细胞的选择性不强。结论两种保种方法保存后弓形虫毒力不变。     

Prevalence of IgG Antibodies to Toxoplasma gondii in Veterinary and Undergraduate Students at Virginia Tech,Blacksburg, Virginia

Toxoplasma gondii is a globally distributed parasitic protozoan that infects humans and other warm‐blooded vertebrates. Felids are the only definitive host for T. gondii, and they excrete oocysts in their faeces. The national prevalence in humans is declining in the United States. This zoonotic organism is of particular interest due to its importance in pregnant women, in individuals with altered immune systems, and in reactivated ocular infections. Exposure to the parasite in humans is usually associated with consumption of raw or undercooked meat or by accidental ingestion of oocysts. It was hypothesized that veterinary students would have a greater chance at exposure to the parasite than an average population of undergraduate students due to increased contact with cats who are infected. A commercially available ELISA was used to examine serum samples from 336 students (252 veterinary students and 84 undergraduate students) at Virginia Polytechnic Institute and State University and the Virginia‐Maryland Regional College of Veterinary Medicine for serum IgG antibodies to T. gondii antigen. The prevalence of T. gondii in these subjects was 5.6% in veterinary school students (n = 252) and 2.4% in undergraduates (n = 84). There was no significant difference (P > 0.05) in the prevalence of T. gondii antibodies in veterinary versus undergraduate students. The overall prevalence of 4.8% in all students in this study reflects the continuing decline of antibodies to T. gondii in humans in the United States.     

Evidence for genetic diversity of Toxoplasma gondii in selected intermediate hosts in Serbia

To contribute to the insight into the worldwide population structure of Toxoplasma gondii, we genetically characterized a total of eight strains isolated from intermediate hosts including humans, sheep and pigeons in Serbia. Although parasite DNA was detected in 28.2% (60/213) of the human samples from 162 patients serologically suspected of active toxoplasmosis, as well as in 5/7 seropositive pigeons and in 2/12 seropositive sheep examined, multilocus PCR-RFLP genotyping, using SAG1, 5′SAG2, 3′SAG2, GRA6, 5′GRA7 and 3′GRA7 as markers, was successful in only four human isolates (of which one was isolated from both the bronchoalveolar lavage fluid and blood samples of a single patient), one sheep and three pigeons. Of the eight isolates, five were type II (62.5%), one was type III, one was atypical, and one had a type I allele at GRA6 as the single locus genotyped. Although type II, as elsewhere in Europe, predominated, these results may suggest a higher genetic diversity of T. gondii in Serbia, reflecting local environmental contamination and also the geographical position of the country in South-East Europe.     

弓形虫基因分型研究进展

弓形虫可感染多种动物引起严重的弓形虫病,为了对不同宿主感染的弓形虫株基因型差异及不同基因型弓形虫虫株的致病力差异进行进一步研究,弓形虫基因型的分析多采用限制性片段长度多态性PCR（PCR-RFLP）、随机扩增多态性DNA、多位点PCR-RFLP分析、DNA序列分析和微卫星DNA序列分析等技术,扩增位点多选择SAG1、SAG2、SAG3、BTUB和GRA6等遗传标记。传统的弓形虫基因型主要有3个,随着基因型研究的不断深入,越来越多的基因型被发现。本文就弓形虫基因分型研究进展进行讨论分析。