摘 要
长时间曝光
运用电镜技术、免疫学技术和分子生物学技术对某实验动物中心发生的小鼠鼠痘进行诊断，为实验小鼠鼠痘的诊断以及制定综合防制措施提供理论依据和参考。
流行病学调查结果表明：因运输和气候变化引起鼠群突然发病，幼龄鼠及繁育高峰种鼠易感染发病，雄性鼠较雌性鼠易感，而老龄鼠不易感。且外观症状不典型，病期极少见小鼠头部、四肢和尾部溃疡。肝、脾、十二指肠和小肠出血、肿胀和坏死等病变较为典型。
动物感染性试验：分别将发病即将死亡的小鼠肝、肺、脾、肾脏等病理组织经碾磨制成细胞悬液，以腹腔注射的方式将病毒接种到BALB／c小鼠和KM小鼠、Wistar 大鼠和家兔体内，接种后第五天，感染小鼠全部死亡。接种后2周，大鼠和兔未表现出异常。结果表明：鼠痘病毒感染并致死BALB/c小鼠和KM小鼠，且BALB/c小鼠感染后的病理变化比KM小鼠更为典型。而鼠痘病毒不感染Wistar大鼠和家兔。
将病死鼠肝、肺、肾、脾和肠等内脏组织，经戊二醛固定、制片，透射电镜观察。结果显示在肝、肺、肾、脾和肠的组织细胞质内均有大量的卵圆形病毒颗粒，其核心呈哑铃状。
将收集的病变肝、脾组织分别放入匀浆器中反复碾磨、离心分离提纯，进行负染后电镜观察，结果观察到大量的病毒颗粒，形状为卵圆形或砖形，其大小为
160-190×250-300nm。
自然感染和人工感染发病的小鼠肝脏、脾脏、肾脏、肺脏、胰、肠等组织常规H. E染色，光镜可观察到自然感染和人工感染鼠痘小鼠主要组织病理变化均表现为脾脏淤血，局灶性坏死和红髓中多核巨细胞显著增多；肝脏局灶性坏死，门管区炎性细胞浸润，出现多核巨细胞；肠表现为出血性坏死性肠炎特征；大脑可见小胶质细胞结节；心脏、肺脏、肾脏和胰脏表现局部的淤血、出血和细胞坏死。
卖马采集发病后47天存活小鼠的血清20份，用ELISA试验方法进行抗体检测，结果阳性率达到100％。
从感染鼠的不同组织中提取病毒核酸，用痘病毒基因组中A－型包涵体蛋白基因作为靶基因，设计合成一对引物，PCR检测技术进行分析，结果表明该DNA片段的核苷酸序列与鼠脱脚病病毒的A－型包涵体蛋白基因序列同源性高达98.6％。
经流行病学、临床症状、病理变化、动物感染试验、电镜观察、组织病理学观察、血清学试验以及PCR技术检测分析进一步鉴定表明，该鼠群发病为急性型流行性鼠痘。
根据鼠痘的流行特点，结合实验室诊断方法，提出鼠痘诊断要点：对于急性型鼠痘建议采取临床病变和电镜检查病原的诊断方法；对于典型鼠痘（亚急性型）应根据临床症状和病理学变化可以做出诊断；对于流行后期的慢性型则采用ELISA方
法测抗体；而对于隐性感染鼠应用PCR检测诊断。
通过对鼠痘特性及诊断方法的一系列研究，提出控制鼠痘发生的综合防疫措施：实验动物规范的引种、运输、检疫及进入；人员的严格准入管理；物品的消毒灭菌进入；实验动物室的卫生防疫消毒管理；灭菌消毒效果的检测；环境微生物的检测和小鼠微生物的检测等，从而保证实验小鼠的质量和动物实验结果的准确性。
关键词： 小鼠 鼠痘 诊断方法 综合防制
Abstract
The moupox in a lab animal center was diagnod by electron microscope, immunology and molecular biology technologies. This will rve as a theoretical basis and refenrence for the diagnosis and integrated control of moupox in mice.
The investigation of epidemiology shows that the moupox was caud by transport and the change of climate. The yong , the breeding boom and the male mice are infected more easily than the old and the female mice. The symptoms of moupox are atypical, the ulcer in mice heads , the four limbs and tails are rarely obrved, but the pathological changes including hemorrhage, swelling and necrosis, etc, in liver, spleen, duodenum and small intestine are typical.
高空作业标准
The experiment of infecting animals: Take the liver, spleen, lung and kindy from the dying mice and grind into cell suspension respectively, then inoculate the BALB/cA mice and KM mice, Wister rats and rabbits by peritoneal injection with the cell suspension. After five days of inoculation, all the infected mice are dead. After two weeks, the rats and rabbits are normal. The results show that the BALB/cA and KM mice can be infected and killed by moupox virus and the pathological changes in BALB/cA are more typical than that in KM mice. Wister rats and rabbits aren’t infected by moupox virus.
The liver, lung, kindy, spleen and intestine of dead mice were fixed with glutaraldehyde and sliced up, a plenty of oval shape viral particles with dumbbell shape core in the cytoplasm were obrved with transmission electron microscope.
Put the liver and spleen into muller and grind repeatly , then parate and purify them by centrifuger, a mount of oval or ellip shape viral particles who size is 160-190×250-300nm were obrved with negative-stain electron microscope .零部件英文
The liver, spleen, kidney, lung, pancreas, intestine of mice infected spontaneously and artificially were dected by HE stain, the main pathological changes include: ecchymoma, focal necrosis and inc
rea of multinuclear giant cell in red pulp in spleen; focal necrosis, infiltration of inflammatory cells in portal area and multinucleate giant cell in liver; hemorrhage and necrosis enteritis in intestine; node of microglial in brain; focal ecchymoma, hemorrhage and cytoclasis in heart, lung, kidneyand pancrea.
The positive rate of antibody detected by ELISA in 20 mice 47 days after infecting is 100%.
孕妇可以喝绿茶吗
The nucleic acid of virus in various tissues of infected mice was extracted and the A-type inclusion body protein of poxviridae virus gene was ud as target gene to compo a pair of primers. The nucleic acid was analized with PCR and the result
showed that the nucleotide quences homology between the virus in infected mice and moupox virus was 98.6%.
Bad on above experiments including epidemiology, clinical symptoms, pathological changes,infection tests, electron microscope, rological experiment and PCR, we can conclude that the dia of this group mice was acute epidemic moupox.
The diagno emphas of moupox by epidemic characters and laboratory tests are as follows: T
he acute moupox was diagnod by the pathological changes of tissues and pathogeny detected by electron microscope, the typical moupox was diagnod by the clinical symptoms and the pathological changes of tissues, the chronic moupox was diagnod by antibody detected with ELISA and the latent infected moupox with PCR. type dia, the methods of diagnosis are changes and finding the pathogeny with; For the typical ratpox(sub-acute type), diagnosing methods are clinical symptoms and pathological changes; For the late epidemy, testing antibody with ELISA; For the latent infected mice, the diagnosing method is PCR.
Bad on a ries of studies on the epidemic charaters and dianosis methods of moupox, an integrated control measure of moupox in mice was made The measures are as follows:the normative introduction, transportion, quarantine and entrance of experimental animals;The strict admittance of personnel;The sterilization of articles;The management of epidemic prevention and disinfection in experimental animal lab;The detection of sterilization effects, microorganisms in environment and mice. So as to ensure the quality of lab mice and veracity of animal experimental result. Key words: mice, moupox, diagnosis methods, integrate control学画漫画基本功
前 言
1.实验动物的现状及应用
实验动物(Laboratory Anima1)是指经人工饲育的、对其携带的微生物实行控制的、遗传背景明确或来源清楚的，用于科学研究、教学、生产、鉴定及其它科学实验的动物(施新猷，2000)，实验动物逐渐广泛地应用于科学实验已有近半个世纪的历史。原来你也在这里吉他谱
随着科学技术的不断发展，实验动物学已逐步形成一门独立的综合性的新兴学科，它不仅仅是对实验动物开发、育种、繁殖和供应等方面的研究，而是对实验动物和动物实验两方面结合起来的研究（陈主初等，2001）。因此，实验动物科学已成为现代科学技术领域不可分割的组成部分，在生物学、医学、药学、兽医学、畜牧学、遗传学、营养学、环境保护、食品卫生、生物制品的生产鉴定以及航天、化工、进口贸易与商品检验等领域的作用日益扩大(郝光荣，2002)。
在生命科学研究领域，实验动物直接关系到人类的身体健康和公共卫生安全。它既是生命科学研究的对象，又作为“活的试剂和度量衡”应用于医药、卫生、科技、教育、环保、化工、农业、商检等行业。目前公认“AEIR” 是进行生命科学研究必需的四个基本条件。所谓“A” 即Animal(实验动物)，“E” 系Equipment(设备)，“I” 为Information(信息)，“R” 是Reagent(试剂)（方喜业等，1995）。这四个基本要素具有同等重要的地位，不能忽略或偏废（李厚达，1992）。而且，各类疾病的诊断、治疗和预防都离不开实验动物，天花的灭绝、人和动物疫苗的研制、异体器官移植以及转基因和克隆技术的重大突破，都是借助于实验动物获得成功（方喜业等，1995）。因此，实验动物作为生命科学发展的基础和重要支撑条件在保障人类健康和优化人类生存条件方面的意义勿庸置疑。
由于实验动物是根据科学研究需要而在实验室条件下有目的、有计划进行人工驯养繁殖和科学培育而成的动物。该动物具有生物学特性明确、遗传背景清楚、表型均一、对刺激敏感和反应一致等特点，这些特点使得仅用少量实验动物就能获得精确可靠的动物实验结果，并具有良好的可重复性（娄成民，2003）。
天谕动漫然而，在应用实验动物进行科学研究过程中，影响动物实验的因素有许多，包括实验动物的环境因素、个体遗传因素、生物因素、营养及其他实验因素等(孙敬方，2001)，在环境因素和动物遗传因素均符合国家标准的情况下，生物因素尤其显得重要。因为，实验动物不同程度的带有细菌、病毒和寄生虫等生物因子，其对实验动物和动物实验的影响是明显的。特别是动物感染细菌、病毒和寄生虫后，显性感染造成动物发病，表现临床症状和病理变化，甚至发生死亡使试验中断，造成人力、物力和时间的极大浪费；而潜在的隐性感染往往无临床体征和症状，常不引
