应用微量血凝及血凝抑制试验诊断非洲狮泛白细胞减少症病例

<正> 猫泛白细胞减少症,又称猫传染性肠炎(猫瘟热),该病病原属于细小病毒科,细小病毒属,猫细小病毒引起猫科动物一种高度接触性急性传染病。主要症状:突然高烧,呕吐、腹泻、脱水及白细胞减少。此病发生于世界许多国家和地区;我国安徽、河南、江苏、山东等地也有此病发生的报道。另外,在西安、齐齐哈尔和杭州某动物园等猫科野生动物也发     

犬细小病毒病诊断技术国家标准

1范围本标准规定了犬细小病毒病的临床检查、血凝与血凝抑制试验、PCR检测的操作方法。本标准适用于犬细小病毒病的鉴定及其流行学调查、诊断和监测。其中病毒分离、血凝与血凝抑制试验、PCR检测适用于犬细小病毒的病原诊断。2试剂和材料2.1FK81细胞(猫肾细胞)。     

伪狂犬病病毒血凝及血凝抑制试验

伪狂犬病毒（pseudorabies virus，PRV）属于疱疹病毒科、疱疹病毒亚科，又称猪疱疹病毒Ⅰ型。某些疱疹病毒，如牛疱疹病毒、马流产病毒、猫鼻气管炎病毒及鸡传染性喉气管炎病毒等都具有凝集动物红细胞的特性。据国外资料报道，PRV也能凝集小鼠红细胞，而目前国内这方面的报道甚少，仅有李学伍等做过此方面的试验研究，本人在李学伍等人试验方法的基础上进行了改进，应用鞣酸法处理小鼠红细胞，提高了实验的敏感性和可重复性，效果十分理想。在猪易感病毒中有很多病毒都能凝集小鼠红细胞，如猪细小病毒，乙型脑炎病毒，冠状病毒等。为了检测伪狂犬病病毒是否能凝集小鼠红细胞及其血凝性可否被特异性抗血清抑制，从而建立一种快速诊断伪狂犬病的方法，特作本试验。     

鸭出血症血凝及血凝抑制试验的建立和应用

鸭出血症是由不同于鸭瘟病毒的鸭新型疱疹病毒引起的鸭的一种传染性疾病。根据该病毒可凝集Balh／c小鼠红细胞的特性，我们建立了鸭出血症血凝及血凝抑制试验，经试验表明该方法是一种快速、实用、特异性强的检测方法。     

血凝与血凝抑制试验4单位病毒抗原配置与标定

为了解决基层兽医实验室检测人员在血凝与血凝抑制（HA和HI）试验中4单位病毒抗原的配置和标定问题的困扰，本试验对GB/T 18936-2020《高致病性禽流感诊断技术》中规定的4单位病毒抗原的配置和标定方法进行了探讨，试验结果表明，通过标定调整的4单位病毒抗原准确性高，从而减少血凝抑制实验的误差。     

马病毒性动脉炎血凝及血凝抑制试验研究与应用

在RK-13细胞上培的马动脉炎病毒在4℃、室温和37℃条件下，用各种动物的红细胞进行血凝试验，证实病毒在不同温度下均可凝集小鼠和鸡的红细胞，但不能凝集牛、绵羊、山羊、马、猪、豚鼠、仓鼠、兔及鹅红细胞，病毒用吐温-80和乙醚处理后，血凝活力会显著增强，其血凝滴度比未经处理的病毒高4-8倍，且血凝像更加清晰，最适处理条件是病毒在冰、浴状态下用终浓度0.06%-0.125%(V/V)二温-80振荡处理15min-60min，再加50%（V/V）乙醚振荡作用5min-15min。用EAV免疫SPF豚鼠，4周后采血做HI和NT试验，显示HI抗体和NT抗体产物成正相关，对550份来自新西兰、吉尔吉斯、内蒙古、新疆的马血清做EAV的抗体检测，比较两个试验，二者符合率为97.8%，血凝抑制抗体与中和抗体效价呈显著的正相关，但抗体效价略低于后者。     

猪细小病毒血凝抑制试验抗原、阳性血清与阴性血清的制备与鉴定

猪细小病毒（Porcine Parvovirus,PPV）血凝抑制试验抗原、阳性血清和阴性血清在猪细小病毒制品的效力检验中不可或缺,对猪细小病毒相关生物制品的质量控制至关重要。试验采用PPV 7909株病毒同步接种PK-15细胞制备血凝抑制试验抗原,用制备的抗原乳化后免疫豚鼠制备阳性血清,同时用未免疫的阴性豚鼠制备阴性血清。对抗原、阳性血清和阴性血清进行鉴定,结果表明,制备的PPV血凝抑制试验抗原HA效价达1:512;阳性血清HI效价达1:1024;阴性血清HI效价＜1:8,且特异性均良好。利用制备的血凝抑制试验抗原、阳性血清与不同PPV疫苗株的灭活抗原、阳性血清进行交叉反应试验,结果表明,制备的抗原与阳性血清具备良好的血清学交叉反应性,可用于PPV制品的统一评价。     

影响血凝试验结果的主要因素

血凝和血凝抑制试验是新城疫、禽流感等疾病的诊断和研究工作中普遍应用的方法。某些病毒（如禽流感病毒和新城疫病毒）能凝集禽类的红细胞,这个现象称为病毒的血凝,可以用来检测样品中病毒的有无和含量的多少,并排特异性反应。因此,血凝和血凝抑制试验可用于鉴定病毒,流行病学调查,检测血清中抗体效价．动物群体疫情的监测等。     

马动脉炎病毒的血凝性研究

在RK－13细胞上培养的马动脉炎病毒（Equine viral arteritis,EAV)在4℃、室温和37℃条件下，用多种动物的红细胞进行血凝试验，病毒在不同温度下均可凝集小鼠和鸡的红细胞，但不能凝集牛、绵羊、山羊、马、猪、豚鼠、仓鼠、兔及鹅的红细胞。将病毒用吐温－80和乙醚处理后，血凝活力会显著增强，其HA效价比未经处理的病毒高4－8倍，且血凝像更加清晰。处理病毒的最适条件是在冰浴状态下用终浓度0．6－1．25mL/L吐温－80振荡处理15－60min，再加1／2体积的乙醚振荡作用5－15min.     

鸡产蛋下降综合症病毒的血凝特性测定

本试验旨在探讨ＥＤＳ病毒血凝反应的最佳反应条件,观察病毒对各种动物红细胞的凝集谱,测定各种因素对病毒血凝素的影响,结果表明,血凝反应宜采用１％的鸡红血球,稀释液用ｐＨ７的生理盐水,室温作用３５ｍｉｎ（分钟）后观察表明,该病毒能凝集鸡,鸭,鹅红细胞,而不能凝集哺乳动物红细胞,耐热,５６℃１ｈ不影响其血凝性,对胰酶和浓度大于０．４％的甲醛敏感;反复冻融８次对血凝效价无影响。     

Comparisons of feline panleukopenia virus, canine parvovirus, raccoon parvovirus, and mink enteritis virus and their pathogenicity for mink and ferrets

Parvoviruses from mink (mink enteritis virus [MEV]), cats (feline panleukopenia virus [FPV]), raccoons (raccoon parvovirus [RPV]), and dogs (canine parvovirus [CPV]) were compared. Restriction enzyme analysis of the viral replicative-form DNA revealed no consistent differences between FPV and RPV isolates, but CPV and MEV isolates could be distinguished readily from other virus types. Feline panleukopenia virus, RPV, and MEV, but not CPV, replicated to high titers in mink. However, on the first passage, disease and microscopic lesions were observed only in mink inoculated with MEV. Feline panleukopenia virus and RPV isolates replicated in ferrets, but disease or microscopic lesions were not observed. Feline panleukopenia virus and RPV isolates could be passaged repeatedly in mink and ferrets. Virulence of FPV and RPV isolates was low compared with that of MEV, and only a single mink inoculated with FPV or with RPV developed clinical disease on the sixth passage of virus.     

猫泛白细胞减少症疫苗的研究进展

猫泛白细胞减少症是一种由猫泛白细胞减少症病毒引起的,猫科动物常见的接触性传染病,临床发病率和死亡率均较高,疫苗免疫接种是预防和控制该病最经济、最直接的措施.然而,国内几无商业化宠物疫苗,特别是猫用疫苗,国外疫苗进口困难,亟需开发新型宠物用高效疫苗.对国内外猫泛白细胞减少症疫苗现状及不同类型疫苗(灭活疫苗、减毒活疫苗等)...     

Feline panleukopenia virus, feline herpesvirus-1, and feline calicivirus antibody responses in seronegative specific pathogen-free cats after a single administration of two different modified live FVRCP vaccines

Two groups of feline panleukopenia virus (FPV), feline calicivirus (FCV), and feline herpesvirus-1 (FHV-1) seronegative cats (five cats per group) were administered one of two modified live feline viral rhinotracheitis, calicivirus, and panleukopenia virus (FVRCP) vaccines and the serological responses to each agent were followed over 28 days. While all cats developed detectable FPV and FCV antibody titers; only two cats developed detectable FHV-1 antibody titers using the criteria described by the testing laboratory. For FPV and FHV-1, there were no differences in seroconversion rates between the cats that were administered the intranasal (IN) FVRCP vaccine and the cats that were administered the parenteral FVRCP vaccine on any day post-inoculation. For FCV, the cats that were administered the IN FVRCP vaccine were more likely to seroconvert on days 10 and 14 when compared to cats that were administered the parenteral FVRCP vaccine.     

用PCR法检测东北虎感染猫细小病毒的研究

根据GenBank中已发表的猫细小病毒基因组中的保守序列 ,利用Goldkey软件设计了一对能扩增 750bp片段大小的引物 ,对某动物园 1只病死东北虎的脾脏样品进行了PCR检测。结果得到了与设计大小完全相符且与标准猫细小病毒扩增产物大小一致的特异核酸带 ,同时对犬瘟热、犬腺病毒、轮状病毒的核酸扩增结果均为阴性。敏感性试验表明 ,此法可检出血凝价为 1 2 8×脾脏匀浆液上清 1 0 - 5倍稀释的模板 ,远高于血凝试验的敏感性 ,为虎、狮、熊猫等野生动物细小病毒病的快速诊断提供了一个有效的方法     

猫泛白细胞减少症病毒的分离与鉴定

从临床表现有体温升高、呕吐、血样腹泻、脱水等症状的疑似猫泛白细胞减少症感染的病例采取粪样28份。从粪便样品中成功分离获得了7株猫泛白细胞减少症病毒（FPV）：JX-1、JX-2、JX-3、JX-4、JX-5、JX-6和JX-7;应用F81细胞增毒,盲传至3代时在F81细胞上产生细胞病变(脱落、变形、游离等);核酸型鉴定证明,FPV毒株的代谢可被5-IUDR所抑制,其核酸属于DNA型;所分离的病毒培养物能凝集猪的红细胞（凝集效价达2⁶～2⁸）,并能被标准FPV阳性血清所抑制;电镜观察病毒粒子外观呈圆形或六边形,直径20～30 nm;该病毒耐酸、耐热、耐乙醚;动物致病性试验,经口感染分离细胞培养毒1 ml,试验组幼猫第7 d发病,采集病猫粪样做HA试验为阳性反应,做HI试验,其凝集猪红细胞的能力被抑制。     

2013年-2015年西安宠物门诊家养猫泛白细胞减少症病例调查

猫泛白细胞减少症(FP)是由猫泛白细胞减少症病毒(FPV)引起的猫科动物的急性高度接触性传染病,临床上常以发热、呕吐、腹泻、脱水、肠炎以及白细胞极度减少为主要特征.本病传播速度快,对猫科动物危害极大.近年来,西安地区家养猫数量不断增加,患FP的病例也随之增多.为了解西安地区家养猫FPV感染和发病情况,为本病的防控提供基础资料,2013年7月~2015年7月,用猫泛白细胞减少症病毒检测试剂盒,对西安市部分宠物(动物)医院接诊的疑似FP病猫的467份粪便进行FPV检测,共检出阳性样品71份,占15.2%(71/467),其中2013年5份,2014年57份,2015年9份;阳性患猫中未免疫接种FPV疫苗的有70份,占98.6%(70/71).确诊病例中47.9%(34/71)临床出现呕吐症状,71.8%(51/71)出现腹泻症状.血液分析发现,84.51%的FPV感染猫的白细胞低于正常值;对71个患猫进行对症和支持治疗后存活率为83.1%(59/71).     

Hemagglutination by canine parvovirus: serologic studies and diagnostic applications

Conditions for canine parvoviral hemagglutination (HA) and hemagglutination-inhibition (HI) reactions were defined. The HA phenomena were used to differentiate canine parvovirus (CPV) from feline panleukopenia virus (FPV), mink enteritis virus (MEV), and minute virus of canines. Serologic comparisons of the CPV, FPV, and MEV by HA-HI and serum-neutralization tests indicated that CPV, FPV, and MEV were antigenically similar but were different from minute virus of canines. Diagnostic application of HA tests to fecal samples from acute cases of enteritis was discussed. Combinating HA tests with HI tests on fecal samples provided a rapid and specific diagnostic method for CPV infection. Secular seroprevalence studies indicated the emergence of CPV infeciton in the United States dog population-at-large in 1978.     

Detection of protective antibody titers against feline panleukopenia virus, feline herpesvirus-1, and feline calicivirus in shelter cats using a point-of-care ELISA

Serum antibody titers are a useful measurement of protection against infection (feline panleukopenia virus [FPV]) or clinical disease (feline herpesvirus-1 [FHV] and feline calicivirus [FCV]), and their determination has been recommended as part of disease outbreak management in animal shelters. The objective of this study was to determine the sensitivity, specificity, and inter-observer and inter-assay agreement of two semi-quantitative point-of-care assays for the detection of protective antibody titers (PAT) against FPV, FHV and FCV in shelter cats. Low sensitivity for FPV antibodies (28%) rendered a canine point-of-care assay inappropriate for use in cats. The feline point-of-care assay also had low sensitivity (49%) and low negative predictive value (74%) for FPV PAT detection, but was highly accurate in the assessment of FHV and FCV PAT. Improvements in accuracy and repeatability of FPV PAT determination could make this tool a valuable component of a disease outbreak response in animal shelters.     

Genetic complexity and multiple infections with more Parvovirus species in naturally infected cats

ABSTRACT: Parvoviruses of carnivores include three closely related autonomous parvoviruses: canine parvovirus (CPV), feline panleukopenia virus (FPV) and mink enteritis virus (MEV). These viruses cause a variety of serious diseases, especially in young patients, since they have a remarkable predilection for replication in rapidly dividing cells. FPV is not the only parvovirus species which infects cats; in addition to MEV, the new variants of canine parvovirus, CPV-2a, 2b and 2c have also penetrated the feline host-range, and they are able to infect and replicate in cats, causing diseases indistinguishable from feline panleukopenia. Furthermore, as cats are susceptible to both CPV-2 and FPV viruses, superinfection and co-infection with multiple parvovirus strains may occur, potentially facilitating recombination and high genetic heterogeneity. In the light of the importance of cats as a potential source of genetic diversity for parvoviruses and, since feline panleukopenia virus has re-emerged as a major cause of mortality in felines, the present study has explored the molecular characteristics of parvovirus strains circulating in cat populations. The most significant findings reported in this study were (a) the detection of mixed infection FPV/CPV with the presence of one parvovirus variant which is a true intermediate between FPV/CPV and (b) the quasispecies cloud size of one CPV sample variant 2c. In conclusion, this study provides new important results about the evolutionary dynamics of CPV infections in cats, showing that CPV has presumably started a new process of readaptation in feline hosts.     

虎源猫瘟热病毒VP2蛋白特异性单克隆抗体的制备

以原核表达的虎源猫瘟热病毒(FPV)VP2蛋白作为免疫抗原,免疫6周龄BALB/c小鼠,用FPV全病毒作为筛选抗原,采用淋巴细胞杂交瘤技术,经过有限稀释法获得了1株可稳定分泌抗虎源FPVVP2蛋白的杂交瘤细胞株3C4。间接ELISA方法测定3C4株单抗腹水效价为1∶12800,亚类鉴定为IgG2a类型,间接免疫荧光试验显示该株单抗能与虎源FPV发生反应,而免疫印迹试验该株单抗未见特异性反应带。