LCMV重组蛋白抗原应用研究

目的用重组蛋白抗原取代淋巴细胞脉络丛脑膜炎病毒(LCMV)抗原用于实验动物感染LCMV检测.方法合成LCMVsRNA第1816～2178位核苷酸序列编码的Np第380～500位氨基酸基因,克隆在Hisx6-GSTpET-28a载体中,表达产物经固定化金属配体亲和层析纯化,建立ELISA检测试剂.结果合成基因表达产物经Ni-NTA-Agarose纯化后纯度达到95%以上.ELISA检测小鼠、地鼠和豚鼠结果与全病毒抗原基本一致,而且特异性较强,操作简便.结论用合成基因表达产物取代LCMV抗原检测病毒抗体,不仅可以消除病毒传播可能,而且特异性强.为实验动物及生物材料质量控制提供安全有效方法.     

上海地区实验动物病原体感染指数分析

目的探索测算感染指数的最佳方法,以便更加科学的反映实验动物的感染状况。方法感染指数,也称感染度,它是对实验动物质量监测的定性指标。通过加工汇总实验动物的病原体感染率,综合反映某一特定实验动物群体的病原体感染状态或趋势的一种指标。结果总体上,小鼠病原体感染指数略有下降,而大鼠的病原体感染指数则逐年上升。分别比较各类病原体感染指数发现,小鼠中寄生虫感染指数较高,而大鼠中细菌感染指数最高。结论感染指数分析揭示了实验动物质量的基本状况,需要加强对小鼠寄生虫的监测,而大鼠的细菌检测需要投入更多关注。小鼠病原体感染情况得到控制,大鼠各类病原体的控制情况均逐年趋于严重。     

实验动物细菌学监测工作中存在的问题及建议

实验动物细菌学质量控制标准是评价实验动物质量的重要指标,本文在分析发达国家实验动物质量标准的基础上,结合目前的检测现状,对我国实验动物细菌学监测标准在检测项目、检测方法、检测频率、取样要求等方面存在的问题进行了分析并提出建议。     

空肠弯曲菌感染动物模型的研究进展

空肠弯曲菌是一种全球关注的人兽共患病原菌,感染后可引起人和动物多种疾病。动物模型是开展致病机理、疫苗评价和药物开发等研究的基础。空肠弯曲菌由于培养条件苛刻以及感染实验动物的疾病相似性、经济性和重复性等因素,仍缺乏良好的感染动物模型,其致病机理迄今尚不清楚。本文对已报道的空肠弯曲菌感染实验动物模型进行综述。     

啮齿动物螺杆菌研究现状

螺杆菌以隐性感染形式普遍存在于大鼠、小鼠、沙鼠等啮齿动物的消化道,可引起各种炎症甚至恶性肿瘤,严重影响实验动物质量,并干扰相关动物实验结果。因此,世界各国及国际实验动物理事会已将该类螺杆菌列为啮齿类实验动物必须排除的病原微生物。本文主要从生物学特性、致病性、检测方法等方面对其研究现状作一简要介绍。     

实验动物福利与人的保护

实验动物学是一门研究实验动物和动物实验的综合性学科。是生命科学研究的基础和支撑条件。随着社会经济的长足发展和生命科学研究的突飞猛进,国际交流与合作的不断加强,实验动物福利倍受人们的关注。进入21世纪后国际上又对从事实验动物饲养管理和动物实验的工作人员的防护问题提出了新的要求,这就要我们必须考虑到人与实验动物如何和谐相处,促进科学发展。既要做到加强动物福利,又要注意人的保护,找到二者之间的平衡点,加快实验动物科学的发展。     

轮状病毒感染成年小鼠的研究

目的研究成年昆明种小鼠对实验感染人轮状病毒(rotavirus,RV)的敏感性。方法在实验条件下,用A组人Wa和恒河猴SA11株RV感染成年昆明种小鼠,观察小鼠的临床反应和排毒情况。结果成年昆明种小鼠感染Wa和SA11RV第二天后出现明显的临床腹泻症状,第四天达到高峰;至少在感染后连续6天的动物大便中可检测到较高滴度的RV抗原。结论成年昆明种小鼠对RV感染有很高的敏感性,可做为动物模型,在RV感染的药物治疗效果评价和疫苗保护性效果评价中具有重要价值。     

建立长爪沙鼠和小白鼠蛲虫病动物模型的可行性研究

本研究选用实验动物长爪沙鼠和小白鼠拟建立新的人蛲虫病动物模型。在实验前,全部动物用复方甲苯咪唑（８０ｍｇ／ｋｇ）驱除肠道线虫。感染动物的蛲虫卵,一种直接取自蛲虫病患儿肛门处,另一种为实验室培养的雌性蛲虫成虫产的卵。实验组动物分别以不同数量的虫卵（１００个／只、４００个／只和１０００个／只）经口感染并分组喂养。同时在小白鼠的实验组中,选一组肌注醋酸可的松。１￣４周后分批解剖动物从肠道收集蛲虫,结果所     

医学实验动物屏障环境内独立通气笼具的安放

目的探讨屏障系统内安放独立通气笼盒（Individually Ventilated Cages,IVC）双保险模式饲养实验啮齿类动物,力求动物饲养、实验的全过程达到SPF级;降低动物实验室硬件建造费和维持费（节能）。方法通过改建一间屏障环境动物实验室,对空气洁净度等相关项目数据按GB14925-2001方法检定,并在该实验室内安放SPC级IVC饲养动物;在超净工作台内做打针投药等动物实验。结果改建的屏障环境动物实验室和IVC,各项检测数据均达到新国标要求。结论医学实验动物屏障环境内安放IVC饲养啮齿类动物、做动物实验,这种双保险模式能全过程达到SPF级要求;饲养、实验人员操作较简便,节能。     

树鼩驯养繁殖实验动物标准及丙型肝炎动物模型技术平台建设的探讨

目前用于生物医学研究的树鼩大多数来源于野生捕获,由于个体间的差异较大,其实验结果的均匀性和可重复性较差,亟待解决树鼩人工饲养设施条件、繁殖关键技术、动物质量标准等一系列基础科学问题。许多文献报道了树鼩与其他实验动物比较,在肝病研究中具有巨大的开发应用潜力,在丙型肝炎动物模型研究中,亟待解决丙型肝炎病毒(HCV)细胞培养体系、HCV病毒感染剂量与感染途径、HCV病毒对树鼩体外、体内感染后病毒核酸检测、抗体检测和病理检测等技术方法的建立及动物模型评价标准问题。本文就上述问题系统性地进行初步报道,旨在为树鼩早日实现实验动物化和丙型肝炎疾病研究中应用提供科学依据。     

Detection of lymphocytic choriomeningitis virus antibody in colonies of laboratory animals in Japan

Indirect fluorescent antibody method was applied for a detection of lymphocytic choriomeningitis virus (LCMV) antibody in colonies of laboratory animals in Japan. The results showed that the antibody exist in SPF mice (3/152, 2.2%) and conventional mice (30/539, 5.6%) with the titers ranging from 1:10 to 1:160. The antibody was also detected in 2.2% (2/89) of Syrian hamsters, and 2.9% (2/68) of Apodemus agrarius, 21.4% (3/14) of Japanese harvest mice which have been maintained as laboratory colony for several years. However, the antibody was not demonstrated in Mongolian gerbils, Suncus murinus, guinea pigs and rats, thus far. These results indicate that LCMV infection is present in laboratory animals in Japan, and pointed out the importance of microbiological monitoring for LCMV.     

Detection of the antibody to lymphocytic choriomeningitis virus in sera of laboratory rodents infected with viruses of laboratory and newly isolated strains by ELISA using purified recombinant nucleoprotein

An enzyme-linked immunosorbent assay (ELISA) was developed to detect the antibody against lymphocytic choriomeningitis virus (LCMV) in sera of laboratory animals. In this ELISA system, LCMV-nucleoprotein (NP) expressed by recombinant baculovirus and purified with high molar urea was used as the antigen. Sera from laboratory animals experimentally infected with the Armstrong strain or the newly isolated M1 strain of LCMV were examined to detect anti-LCMV antibody by the ELISA system, and the reactivity was compared with that of IFA test. Regardless of LCMV strain, all the sera of adult mice infected with LCMV were positive with very high optical density (OD). Also, the sera from mice neonatally infected with LCMV M1 strain were positive with slightly lower OD than adult mice. In contrast, all the sera of uninfected mice were negative to LCMV-NP antigen. Similarly, anti-LCMV antibodies were detected in all the sera of hamsters, mastomyses, and gerbils infected with the LCMV Armstrong strain. The results of the ELISA were in complete agreement with those of IFA, and indicate the high sensitivity and specificity of the ELISA system in the detection of anti-LCMV antibody. Because this ELISA system does not require handling infectious LCMV in the course of the antigen preparation and serological assay, there is no risk of contamination in the laboratory or nearby animal facility. In addition, by using negative control antigen in parallel with positive antigen in ELISA, we can exactly check the LCMV contamination in laboratory animals.     

五种国产与进口小鼠病毒ELISA抗体检测试剂盒的比较研究

目的评价国产小鼠病毒抗体ELISA检测试剂盒。方法选择国产与进口小鼠淋巴细胞脉络丛脑膜炎病毒（LCMV）、肝炎病毒（MHV）、仙台病毒（SV）、腺病毒（MAV）、细小病毒（MPV）ELISA抗体检测试剂盒,进行敏感性、特异性、精密性、稳定性、可信度试验比较。结果国产与进口试剂盒：同种试剂盒之间灵敏度相差最低为2倍,差异显著（P〈0.05）,最高为16倍,差异极显著（P〈0.01）;特异性试验显示每种试剂盒,与其他4种病毒均无交叉反应;精密性试验显示5种试剂盒批内平均变异系数均小于10%;稳定性试验显示5种试剂盒相对偏差均小于25%;分别选择已知36份小鼠血清进行检测,国产和进口LCMV、MHV、SV、MPV符合率均为100%;国产MAV符合率为86.1%,进口MAV符合率均为100%,二者之间差异极显著（P〈0.01）。结论除国产MAV试剂盒敏感性、可信度低于进口外,国产LCMV、MHV、SV、MPV试剂盒与进口同种试剂盒相比,在敏感性、特异性、精密性、稳定性和可信度方面均良好。     

Contamination of transplantable murine tumors with lymphocytic choriomeningitis virus

Lymphocytic choriomeningitis virus (LCMV) was isolated from a transplantable tumor after mice bearing the tumor began to die prematurely. Tumor lines, mice and laboratory personnel that had an association with the index laboratory were tested for LCMV infection. Testing of tumor lines from the index laboratory and four other laboratories revealed that 16 of 55 tumor samples used in vivo and one of eight tumor samples maintained in vitro were contaminated with LCMV. Laboratory personnel and uninoculated mice that were exposed to infected tumors had no LCMV antibody. The use of carefully monitored seed stocks is recommended to protect transplantable tumors that may be inadvertently contaminated by viruses.     

玩具对小鼠生长发育及繁殖性能的影响

目的：探讨玩具对实验小鼠生长发育及繁殖性能的影响。方法选取7-8周龄ICR小鼠,在生产过程中加入玩具进行干预,观测其对小鼠生长发育及繁殖性能方面指标的影响。结果除产仔数外,实验组各项繁殖性能指标均优于对照组,且差异显著（ P ＜0.05）。并且随干预时间延长,差异的显著性有进一步增加的趋势。实验组小鼠的生长发育水平相比于对照组有明显变化,且表现出有随干预时间延长而产生差异的趋势,但目前尚无统计意义（ P ＞0.05）。结论玩具对ICR小鼠的生长发育及繁殖性能有一定程度的积极影响,在小鼠的生产和饲育过程中值得借鉴。     

两种方法对上海及周边地区实验大小鼠螺杆菌携带情况的调查

目的了解上海及周边地区实验小鼠、大鼠螺杆菌携带情况,为我国实验动物等级及监测标准的制定提供参考和依据。方法PCR法共检测了352只小鼠（清洁级101只,SPF级251只）,101只大鼠（清洁级69只,SPF级32只）;ELISA法共检测了88只小鼠（清洁级26只,SPF级62只）,165只大鼠（清洁级84只,SPF级81只）;并对其中88只小鼠、101只大鼠的PCR和ELISA法阳性检测率进行比较。结果PCR法检测小鼠平均阳性率为35．8％（126／352）,清洁级阳性率为51．5％（52／101）,SPF级阳性率为29．5％（74／251）;大鼠平均阳性率为70．3％（71／101）,清洁级阳性率为69．6％（48／69）,SPF级阳性率为71．9％（23／32）;ELISA法检测小鼠平均阳性率为15．9％（14／88）,清洁级阳性率为19．2％（5／26）,SPF级阳性率为14．5％（9／62）;大鼠平均阳性率为52．7％（87／165）,清洁级53．6％（45／84）,SPF级51．9％（42／81）;88只小鼠PCR法阳性检测率为72．7％（64／88）,ELISA法阳性检测率为15．9％（14／88）;101只大鼠PCR法阳性检测率为70．3％（71／101）,ELISA法阳性检测率为49．5％（50／101）。结论上海及周边地区实验大鼠、小鼠中皆存在着不同程度的螺杆菌感染,两种方法阳性检出率比较结果表明回盲部内容物PCR法较检测血清中抗螺杆菌抗体ELISA法更为敏感。     

Serological survey of virus infection among wild house mice (Mus domesticus) in the UK

The serological prevalence of 13 murine viruses was surveyed among 103 wild-caught and 51 captive-bred house mice (Mus domesticus), originating from several trapping locations in northwest England, using blood samples obtained during routine health screening of an established wild mouse colony. A high proportion of recently caught wild mice were seropositive for mouse hepatitis virus (86%), mouse cytomegalovirus (79%), mouse thymic virus (78%), mouse adenovirus (68%), mouse parvovirus (59%) and minute virus of mice (41%). Seroprevalences of lymphocytic choriomeningitis virus (LCMV), orthopoxvirus, reovirus-3 and murid herpesvirus 4 (MuHV-4, also called murine gamma-herpesvirus [MHV-68]) were low (3-13%), and no animals were seropositive to Sendai virus, pneumonia virus or polyomavirus. Seroprevalence in wild-caught animals that had been in captivity for over six months was generally consistent with the range found in recently caught wild animals, while seroprevalence was generally much lower in captive-bred mice despite no attempt to prevent viral spread. A notable exception to this was LCMV, which appeared to have spread efficiently through the captive population (both captive-bred and wild-caught animals). Given the known viral life cycles in laboratory mice, it appears that viral persistence in the host was an important contributing factor in the spread of infection in captivity.     

稀有鮈鲫对重铬酸钾和3,4-二氯苯胺急性毒性研究

目的和方法稀有鮈鲫(Gobiocypris rarus)作为一种本土特有的小型鱼类,是我国正在标准化的化学品毒性测试生物之一。为评价稀有鮈鲫鱼类急性毒性实验中效应的稳定性和可重复性,研究优化筛选了重铬酸钾和3,4-二氯苯胺两种化学品,分别在单一实验室内及不同实验室间进行急性毒性验证实验。结果对于来源相同、体长固定的稀有鮈鲫,无论在实验室内还是实验室间,两组化学品的96 h LC50值均在x珋±2s的质量控制范围之内,所有结果体现了很好的稳定性和可重复性。结论根据研究获得的数据可建立一个有效的稳定性和可重复性衡量数据库,用来评估之后其他单个实验的可靠性;稀有鮈鲫作为一种具有潜力的生态毒性测试的生物种类,具有成为标准实验动物的潜能。