犬科、猫科动物细小病毒血清抗体调查

肉食兽细小病毒属于细小病毒科、细小病毒属中的一类病毒,能够感染多种动物,导致犬的出血性肠炎、幼龄犬的心肌炎、猫的白细胞减少、出血性肠炎、幼龄猫的共济失调症以及水貂的肠炎等多种疾病.血清学调查发现,由肉食兽细小病毒引起的疾病存在于世界各地.在我国,无论是家养还是野生动物均有细小病毒相关疫病的流行,对我国犬科和猫科动物的生存和健康构成巨大威胁(许树林等,1996;宋桂强等,2007).     

四种动物病毒的细胞培养及血凝检测的比较研究

四种动物病毒的细胞培养及血凝检测的比较研究李天宪,赵林,罗怡珊,冯锋（中国科学院武汉病毒研究所,武汉４３００７１）关键词细小病毒,细胞培养,细胞病变,血凝试验云豹肠炎病毒（ＬＰＶ）、水貂肠炎病毒（ＭＥＶ）、犬肠炎病毒（ＣＰＶ）和猫泛白细胞减少症病毒（...     

犬细小病毒单克隆抗体的制备及其与免疫血清在血凝抑制中的比较

将用犬细小病毒免疫的ＢＡＬＢ／ｃ小鼠的脾细胞与Ｓｐ２／０骨髓瘤细胞在聚乙二醇作用下融合,经筛选、克隆,得到６株稳定分泌犬细小病毒单克隆抗体的杂交瘤。用杂交瘤腹水作血凝抑制试验,检测分别含有犬细小病毒、猫泛白细胞减少症病毒和水貂肠炎病毒的标本,及ＣＰＶ感染犬粪便标本,并与免疫血清作对比。结果表明,用单克隆抗体作血凝抑制试验,比用免疫血清作具有特异性高、操作简便省时等优点,而二者敏感性一致。     

犬细小病毒:从起源到进化

犬细小病毒(Canine parvovirus,CPV-2)首次分离于1978年,被认为是由遗传关系相近的猫泛白细胞减少症病毒(FPLV)或其他肉食兽细小病毒(FPLV-like virus)跨宿主感染犬产生的新病原.其感染能引起新生犬急性心肌炎或幼犬出血性肠炎,造成较高的发病率和死亡率.该病原爆发后短短一年时间内即广泛流行到世界各地.随着CPV-2对宿主的适应和变异,新抗原变异型(CPV-2a、CPV-2b和CPV-2e)不断产生并在世界各地逐步替代了CPV-2的流行.伴随CPV-2抗原变异的同时,其对宿主(犬、猫)嗜性、毒力等生物学特性也随之改变.本文综述了CPV-2过去30多年在世界流行和变异情况,并探讨了基因变异在病毒跨宿主传播中的意义.     

犬细小病毒的PCR诊断试剂盒的研制

目的　建立检测犬细小病毒的PCR诊断试剂盒。方法　通过在犬细小病毒 (CPV)的基因组中设计的特异性寡核苷酸引物 ,利用PCR技术研制犬细小病毒的PCR诊断试剂盒。结果　在特定的反应条件下 ,使用该试剂盒能特异地扩增含有犬细小病毒的样品 ,且能检出痕量 (10ng)的犬细小病毒DNA ,被测粪样只需进行简单煮沸就能进行PCR反应 ,该试剂盒能在常温下保存 1周以上、4℃保存 1年以上。同血凝试验 (HA〕及单克隆抗体ELISA方法比较 ,对 6 6份样品进行检测 ,显示该PCR试剂盒具有特异、灵敏等优点。结论　成功研制了犬细小病毒的PCR诊断试剂盒 ,利用该试剂盒能从DNA水平上对犬细小病毒进行监控     

猫肾F_(81)传代细胞中犬细小病毒原位PCR检测方法

目的　建立在猫肾F81 细胞中犬细小病毒原位PCR的检测方法。方法　在猫肾F81 细胞上感染犬细小病毒 ,设计特异性引物 ,用直接原位PCR法在染毒 12h ,2 4h ,48h细胞片上检测出犬细小病毒 ,并与常规免疫组化的方法进行了比较。结果　在染毒 48h的细胞片上 ,用阳性记分法将两种检测方法得到的阳性细胞进行统计比较 ,差异极显著 (P <0 .0 0 1) ,用原位PCR法所得出的阳性率高。结论　原位PCR法检测犬细小病毒具有敏感性高和组织定位的优点     

猫肾F81传代细胞中犬细小病毒原位PCR检测方法

目的建立在猫肾F81细胞中犬细小病毒原位PCR的检测方法.方法在猫肾F81细胞上感染犬细小病毒,设计特异性引物,用直接原位PCR法在染毒12h,24h,48h细胞片上检测出犬细小病毒,并与常规免疫组化的方法进行了比较.结果在染毒48h的细胞片上,用阳性记分法将两种检测方法得到的阳性细胞进行统计比较,差异极显著(P＜0.001),用原位PCR法所得出的阳性率高.结论原位PCR法检测犬细小病毒具有敏感性高和组织定位的优点.     

半嵌套式PCR技术区别检测犬细小病毒疫苗株和强毒株的研究

犬细小病毒病是由犬细小病毒(Canineparvovirus,CPV)引起的一种多发于幼犬的致死性传染病,主要表现为出血性肠炎和心肌炎[1].在自然条件下本病多呈散发,而在养犬比较集中的地方则常群发[2].     

水貂病毒性肠炎的防制研究——Ⅰ.病毒的分离和鉴定

从山东和青海省的水貂肠炎流行区采集的64份病料中,用幼猫肾次代细胞分离到6株水貂肠炎病毒(MEV)。经电镜、理化学和血清学试验证实为水貂细小病毒(Mink Parvovirus)。     

犬细小病毒诊断与防制研究进展

犬细小病毒是严重危害野生动物驯养繁殖产业的重要病毒性传染病之一,具有感染高、发病急、病程短和死亡率高等特点,且随着病毒的不断变异,其宿主范围不断扩大,除感染狐狸、犬、狼等犬科动物外,还能感染猫、豹、熊、虎和果子狸等其它多种动物,对我国的野生动物保护、驯养繁殖和疫源疫病监测工作的影响越来越大。本文对犬细小病毒的诊断及其防制进行了概述,以期为野生动物感染犬细小病毒的防控提供科学依据。     

Autometallography: tissue metals demonstrated by a silver enhancement kit

In biological tissue, minute accumulations of gold, silver, mercury and zinc can be visualized by a technique whereby metallic silver is precipitated on tiny accumulations of the two noble metals, or on selenites or sulphides of all four metals. In the present study a silver enhancement kit, primarily intended for the amplification of colloidal gold particles, has been used to demonstrate these catalytic tissue metals. Sections from animals exposed intravitally to aurothiomalatate, silver lactate, mercury chloride, sodium selenite or perfused with sodium sulphide were subjected to a commercial silver enhancement kit (IntenSE, Janssen Pharmaceutica). It was found that the kit performs adequately to the silver lactate gum arabic developer and to the photographic emulsion technique. The kit can be used as a silver enhancement medium for the demonstration of zinc by the Neo-Timm and selenium methods and for demonstration of gold, silver, and mercury in tissues from animals intravitally exposed to these metals. It can also be used for counterstaining silver treated osmium fixed tissues embedded in plastic.     

HIV-1 p24和SIV p27两种ELISA试剂盒检测SIV p27抗原的比较

目的由于检测SIV p27抗原试剂盒来源困难,有时不稳定,鉴于HIV-1 p24与SIVp27有较强的交叉抗原,本研究比较HIV-1 p24和SIV p27两种ELISA试剂盒检测SIV p27抗原得出的结果是否存在一定的相关性。方法 HIV-1 p24和SIV p27两种ELISA试剂盒定性和定量检测样品中SIV p27抗原,并对检测结果进行回归和相关分析。结果 HIV-1 p24和SIV p27两种ELISA试剂盒检测SIV p27抗原的灵敏度分别是150 pg/mL和62.5 pg/mL。两种试剂盒检测病毒液和血浆中SIV p27抗原的定性结果一致。定量结果的统计分析得出病毒液的直线回归决定系数R2=0.857,直线相关系数r=0.926,P〈0.01,直线正相关程度较高;血浆的直线回归决定系数R2=0.512,直线相关系数r=0.716,P〈0.05,直线正相关程度较低。结论 HIV-1 p24 ELISA试剂盒能够替代SIVp27 ELISA试剂盒定性检测病毒液和血浆中SIV p27抗原,但只能定量检测病毒液中SIV p27抗原。     

Quality Evaluation of Methyl Binding Domain Based Kits for Enrichment DNA-Methylation Sequencing

DNA-methylation is an important epigenetic feature in health and disease. Methylated sequence capturing by Methyl Binding Domain (MBD) based enrichment followed by second-generation sequencing provides the best combination of sensitivity and cost-efficiency for genome-wide DNA-methylation profiling. However, existing implementations are numerous, and quality control and optimization require expensive external validation. Therefore, this study has two aims: 1) to identify a best performing kit for MBD-based enrichment using independent validation data, and 2) to evaluate whether quality evaluation can also be performed solely based on the characteristics of the generated sequences. Five commercially available kits for MBD enrichment were combined with Illumina GAIIx sequencing for three cell lines (HCT15, DU145, PC3). Reduced representation bisulfite sequencing data (all three cell lines) and publicly available Illumina Infinium BeadChip data (DU145 and PC3) were used for benchmarking. Consistent large-scale differences in yield, sensitivity and specificity between the different kits could be identified, with Diagenode''s MethylCap kit as overall best performing kit under the tested conditions. This kit could also be identified with the Fragment CpG-plot, which summarizes the CpG content of the captured fragments, implying that the latter can be used as a tool to monitor data quality. In conclusion, there are major quality differences between kits for MBD-based capturing of methylated DNA, with the MethylCap kit performing best under the used settings. The Fragment CpG-plot is able to monitor data quality based on inherent sequence data characteristics, and is therefore a cost-efficient tool for experimental optimization, but also to monitor quality throughout routine applications.     

S79株腮腺炎病毒制备纯化抗原用于诊断试剂的研究

以S79株腮腺炎病毒制备纯抗原,用于制备检测腮腺炎抗体(IgG)的ELISA试剂盒。将腮腺炎病毒S79株培养液用中空纤维超滤器进行浓缩,经PEG沉淀后,采用蔗糖密度梯度离心法纯化抗原;以纯化的S79株腮腺炎病毒为包被抗原,制备成ELISA腮腺炎病毒抗体(IgG)诊断试剂,并与SIGMA同类产品进行比较。结果显示,经纯化的S79株抗原的比活力为412.9HAU/mg,杂蛋白清除率为99.5%。应用纯化的S79株腮腺炎病毒抗原制备的ELISA试剂,与SIGMA试剂比较,敏感度为94.4%,特异度为91.7%,一致性为93.3%。结论,以纯化的S79株腮腺炎病毒为包被抗原,可用于大批量ELISA腮腺炎抗体诊断试剂盒的制备。     

小鼠肝炎病毒抗体检测试剂盒的标准化研究

目的筛选适合制备检测试剂盒的抗原组合。方法用 ＥＬＩＳＡ方法,将５株小鼠肝炎病毒（ＭＨＶ）抗原分别组合作为包被抗原,研究ＭＨＶ抗原不同组合与标准毒株抗体的反应性,分析ＭＨＶ各毒株间的差别,进一步比较不同组台抗原的灵敏性、特异性。结果通过对１９９７～１９９９年送检２３８只普通级实验小鼠检测,结果表明,ＭＨＶ的感染率虽有下降趋势,但感染率仍然很高（５９％～８７％）。结论我国分离的ＭＨＶ－ＨＵ７９株,在本次研究中表明能够用于ＭＨＶ抗体检测试剂盒。     

猴T淋巴细胞趋向性病毒1型双抗原夹心ELISA检测试剂盒的研制

目的研制灵敏度和特异性高的检测实验猴血清中T淋巴细胞趋向性病毒-1型（STLV-1型）／E体的双抗原夹心ELISA（dsELISA）检测试剂盒。方法采用经原核表达系统表达并纯化的人T淋巴细胞白血病病毒-1型（HTLV-1型）的Env蛋白作为包被用抗原,建立了检测STLV-1的dsELISA诊断方法。通过优化反应条件和筛选试剂,确定了dsELISA诊断试剂盒的相关条件,并经敏感性、特异性和重复性试验考查该试剂盒质量。结果试剂盒特异性好,批内重复试验变异系数（CV）〈7％,批间重复试验CV〈10％。对200份猴血清进行随机检测,与国际公认的诊断试剂盒（美国BioReliance公司）的符合率为97％。结论本试剂盒可初步应用于临床上实验猴STLV-1型抗体的检测。     

改良幽门螺杆菌抗原检测试剂盒 检测粪便幽门螺杆菌抗原的临床评价

目的评估改良幽门螺杆菌抗原检测试剂盒检测粪便幽门螺杆菌抗原(Helicobacter pylori stool antigen,HpSA)的准确性以及临床应用价值。方法采用随机、双盲、双验证和与~(13) C呼气试验(~(13) C-UBT)对比的方法,对门诊175例接受~(13) C-UBT检测的患者,采用最新研制出的一种改良幽门螺杆菌抗原检测试剂盒(胶体金法)检测粪便幽门螺杆菌抗原,以~(13) C-UBT检测结果为诊断H.pylori感染的"金标准",并将两者进行对比研究,所有检测结果均拍照存档,采用随机、双盲和双验证法,以期客观真实地评价改良幽门螺杆菌抗原检测试剂盒检测粪便幽门螺杆菌的效果。结果改良幽门螺杆菌抗原检测试剂盒检测HpSA敏感度为90.48%,特异度为90.00%,Youden指数为80.48%,Kappa值为0.799;HpSA检测ROC曲线下面积为0.902±0.027,与完全无诊断价值的机会线下面积0.50相比,差异有统计学意义(P=0.000);Spearman相关系数r=0.800,P=0.000。结论改良幽门螺杆菌抗原检测试剂盒能准确检测H.pylori感染,其操作简便,可作为非侵入性诊断H.pylori感染筛查以及流行病调查的一种方法,将来有望成为患者家庭自查幽门螺杆菌的一种方法。     

Usefulness of the diagnostic kit Enteroplast for identification of Enterobacteriaceae

Comparative evaluation of biochemical properties determined by ENTEROPLAST kit (P.P.Z. Plastomed) and biochemical set of tests (conventional method) of 140 strains representing 12 genera of Enterobacteriaceae family was undertaken. Consistent results positive or negative (in 12 biochemical tests) were obtained in 93%. The highest percentage of inconsistent results of biochemical reactions (positive in conventional method and negative in ENTEROPLAST kit) were observed in following tests: growth on Simmons medium--17.8%, MR--7.8%, fermentation of raffinose--7.2%. Significantly lower percentage of inconsistent results was found in the case of false positive reactions (negative in conventional method and positive in ENTEROPLAST kit). In summary, it seems that Enteroplast kit can be used for routine diagnostic examinations for identification of rods of Enterobacteriaceae family, basing on their biochemical properties.     

Doctor on a mountaineering expedition.

Doctors are welcome members on mountaineering expeditions to remote areas, but practical advice on how to prepare and what kit to take can be difficult to find. This article is a ragbag of useful advice on diverse topics. It explains the necessary preparation, provides tips for a healthy expedition, and summarises the common disorders encountered at high altitude. The comprehensive drug and equipment lists and first aid kit for climbers were used for the 1992 Everest in winter expedition. They are there to be sacrificed to personal preference and the experience and size of individual expeditions.