猪瘟抗体检测技术的研究现状及发展趋势

简要叙述了FAVN、NPLA、ELISA、兔体中和等猪瘟抗体检测技术的原理及其应用现状,并通过对猪瘟抗体检测技术发展趋势的介绍,显示了猪瘟参考血清等相关标准物质在猪瘟抗体检测技术应用与发展中的价值及其今后的发展前景.     

猪瘟抗体的几种常用检测方法概述

近年来,我国猪瘟免疫失败常有发生,给养猪业带来较大的经济损失。猪瘟抗体检测能准确掌握猪群的猪瘟抗体水平,对猪瘟疫苗免疫效果评价、猪瘟免疫程序制定、猪瘟流行状态监测均具有重要意义。文章就猪瘟抗体中和试验、间接血凝试验、酶联免疫吸附试验、胶体金免疫技术等常用检测方法的研究与应用现状及其优缺点进行全面论述,为各级兽医部门及养殖户寻找和建立适合自身的抗体检测方法提供参考。     

两种不同检测法对猪瘟抗体阴性猪筛检的比较

猪瘟抗体阴性猪是猪瘟活疫苗安检的重要实验动物,目前对于猪瘟抗体阴性猪筛选是采用兔体中和反应法,实验复杂,工作量大并且易受实验兔个体差异等因素影响。随着生物学及其技术的发展,ELISA被广泛用于生物制品的各个领域。本试验应用ELISA法和兔体中和反应法对猪瘟抗体阴性猪进行筛选,共检测160份血清,检测结果显示两种方法结果有很高的一致性,但ELISA法更为敏感,快捷,更适用于猪瘟抗体阴性猪的筛选。     

猪瘟胶体金免疫快速检测技术研究进展

猪瘟是由猪瘟病毒引起的一种严重危害养猪业的毁灭性传染病。胶体金免疫技术是一种新型免疫学检测技术,具有操作简便、快速、灵敏度高、特异性强、无需特殊仪器等优点,在猪瘟的快速诊断中得到了广泛应用,显示出了良好的应用前景。本文概述了胶体金免疫技术在猪瘟抗原及抗体快速检测上的应用研究进展,以期为猪瘟的有效防控提供参考。     

猪瘟胶体金免疫快速检测技术研究进展

猪瘟是由猪瘟病毒引起的一种严重危害养猪业的毁灭性传染病。胶体金免疫技术是一种新型免疫学检测技术,具有操作简便、快速、灵敏度高、特异性强、无需特殊仪器等优点,在猪瘟的快速诊断中得到了广泛应用,显示出了良好的应用前景。通过概述胶体金免疫技术在猪瘟抗原及抗体快速检测上的应用研究进展,对现状及发展前景作一简要综述,以期为猪瘟的有效防控提供参考。     

非洲猪瘟病毒及诊断技术研究进展

非洲猪瘟病毒是非洲猪瘟病毒科、非洲猪瘟病毒属的唯一成员,病毒基因组庞大,能编码100种以上的蛋白质分子。20世纪20年代该病毒首次在非洲的肯尼亚被发现,经过几十年,已经在世界多个国家不断蔓延,我国于2018年8月首次发生非洲猪瘟疫情。目前,非洲猪瘟抗原检测方法有红细胞吸附试验、荧光抗体试验、PCR技术、荧光定量PCR技术、环介导等温扩增技术等。抗体检测技术主要是酶联免疫吸附试验、胶体金试纸条法等。根据非洲猪瘟疫情在我国的发生情况,中国动物疫病预防控制中心组织了非洲猪瘟现场快速检测评审,对各种检测方法进行了技术把关,为非洲猪瘟病毒的检测和防控提供了技术保证。     

乳猪猪瘟被动与主动免疫效果的监测报告

为了摸清猪瘟母源抗体的作用情况 ,了解乳猪实施接种猪瘟三联苗后产生免疫力的确切效果 ,笔者于 2 0 0 0年 3～ 6月对我州坝固种畜场的 4头孕猪 (大约克 )及其仔猪 (与长白杂交的后代共 36头 )进行了被动与主动免疫效果的监测。应用正向血凝试验检测生产母猪及其仔猪血清中的抗猪瘟抗体滴度 ,同时还检测实施主动免疫后乳猪不同时期血清中抗猪瘟抗体的消涨变化。发现孕猪有抗猪瘟抗体的比率为75 % ;其仔猪通过母体获得抗猪瘟抗体的比率为 41 67% ;对未满双月的仔猪接种猪瘟三联苗后 ,其血清中产生有效抗猪瘟抗体的比率比通过母体被动免疫而获…     

猪瘟胶体金快速诊断法的研究进展

胶体金免疫层析技术是一种新型的免疫检测技术,具有操作简单、方便快速、特异性强、灵敏度高、无需特殊仪器等优点。本文简要阐述了胶体金技术在检测猪瘟抗原及猪瘟抗体上的应用现状,并提出了该技术目前存在的问题及未来的发展方向。     

猪瘟实验室抗体检测技术

在猪瘟的确诊、预防、流行病的专研、血清的调查和疫苗的开发中,猪瘟抗体测试技术被广泛使用,但每种检测技术有各自的利弊,本文介绍了多种猪瘟抗体检测技术,读者可以根据实际情况确定相关的测试技术。     

猪瘟胶体金快速诊断法的研究进展

胶体金免疫层析技术是一种新型的免疫检测技术,具有操作简单、方便快速、特异性强、灵敏度高、无需特殊仪器等优点。本文简要阐述了胶体金技术在检测猪瘟抗原及猪瘟抗体上的应用现状,并提出了该技术目前存在的问题及未来的发展方向。     

Reference intervals for feline cerebrospinal fluid: biochemical and serologic variables, IgG concentration, and electrophoretic fractionation

Reference intervals are reported for feline CSF biochemical and serologic variables, IgG concentration, and electrophoretic fractionation, derived from 58 clinically normal adult cats that did not have histologic lesions of the CNS. There was no apparent effect of age on any variable. The CSF total protein concentration was significantly (P = 0.012) greater in males than in females, but all other variables were unaffected by gender. The only variable that had a statistically significant correlation with its corresponding blood concentration was IgG. Blood contamination of the CSF affected the following CSF variables: total protein concentration, activities of lactate dehydrogenase and creatine kinase, IgG ratio, and gamma-globulin percentage. The reference intervals proposed for feline CSF were derived from 33 cats with CSF RBC count less than 31 cells/microliters. Reference limits for CSF with 31 to 1,700 RBC/microliters also are reported.     

Reference intervals for feline cerebrospinal fluid: cell counts and cytologic features

Reference intervals for feline CSF cell counts and cytologic variables were determined. Values were derived from 58 adult cats that had normal neurologic examination findings and did not have histologic lesions of the CNS. Effect of age or gender was not apparent for any CSF variable, and no CSF variable was significantly correlated with its corresponding blood value. Total WBC count and neutrophil and eosinophil percentages were positively correlated with the CSF RBC count. Thus, proposed reference intervals for feline CSF were derived from 33 cats with CSF RBC count of less than 31 cells/ul. Data for CSF samples with range between 31 and 1,700 RBC/microliters were also determined. Erythrocyte count was not significantly different in CSF collected, using 20- or 22-gauge spinal needles.     

Classical swine fever (CSF): a historical review of research and vaccine production on the Isle of Riems

A review on classical swine fever (CSF) research and vaccine production is given about four historical periods (1924-1948, 1949-1969, 1970-1991, since 1992). Similar as to research on foot and mouth disease, applied topics as diagnosis, pathogenesis, epidemiology and control represented the CSF research over many years. The development of vaccines and application procedures, e.g. oral and aerogenic immunisation and combined vaccines for large pig farms were the prominent investigations between 1950 and the middle of 1980s. After being reduced in the first years after affiliation to the Federal Research Centre for Virus Diseases of Animals, CSF is one of the main topics of the research on the Isle of Riems, not at least because nowadays the German National Reference Laboratory for CSF was established on the Island.     

ELISA法检测猪瘟抗体消长规律的试验研究

本试验采用间接ELISA方法,检测猪瘟(CSF)常规免疫仔猪的CSF抗体水平。试验分A、B两组,A组首免注射4头份猪瘟兔化弱毒疫苗,B组首免注射2头份猪瘟兔化弱毒疫苗;二免两组均注射2头份猪瘟兔化弱毒疫苗。试验结果表明,注射疫苗后10d,开始产生抗体,20d-30d抗体达到峰值,二免后抗体峰值维持40d以上。首免4头份的仔猪比首免2头份的仔猪CSF抗体峰值出现的早,维持时间长。     

猪瘟猪伪狂犬病二联苗对仔猪免疫效果观察

本文探讨了猪瘟(CSF)、猪伪狂犬病(PR)活苗不同时间对仔猪免疫效果的相互影响。结果表明:同时免疫两种活疫苗(二联苗组),PR会干扰CSF前期抗体的产生。二联苗组猪瘟抗体3周后才达到阳性,而猪伪狂犬病抗体在7日时就达到阳性,两种抗体均在49日达到峰值,峰值均比单一免疫、分时免疫的峰值略低。分时免疫两种疫苗产生的抗体均比单一免疫时的抗体要低,且相互干扰不明显。     

Genetic typing of classical swine fever viruses--a review

Classical swine fever (CSF) is a notifiable disease of domestic pigs and wild boar. It is caused by the highly contagious CSF virus and in its acute form the disease generally results in high morbidity and mortality. Due to the great economical impact an outbreak can cause to the pig industry it is one of the most important swine diseases worldwide. To limit the damage in the case of a new outbreak it is necessary to identify the virus as fast as possible. This information helps epidemiologists to trace the origin of the virus and to follow the virus spread. Genetic typing revealed that CSF virus genotypes, subgroups and types show a regional distribution making it an important tool for epidemiologists. Meanwhile, besides epidemiological data and nucleotide sequences from European isolates, information from isolates from South- and Central America, the Caribbean, Asia and recently from South Africa have become available. The data are stored in a database in the EU Reference Laboratory for CSF, accessible by the WWW (http://viro08.tiho-hanno ver.de). A new module was implemented that allows efficient automated genotyping.     

Composition and analysis of cerebrospinal fluid in clinically normal adult cattle.

Cerebrospinal fluid and serum were obtained from 16 clinically normal adult cows (11 dairy, 5 beef). Sodium, potassium, magnesium, total protein, and albumin concentrations, osmolality, and lactate dehydrogenase and creatine kinase activities, were quantified in CSF and serum. Total and differential cell counting, protein electrophoresis, and IgG quantification were performed on CSF. Statistical analyses of these variables, including mean, SEM, range, and 95% confidence intervals, were performed. Effects of blood contamination were evaluated, and were found to be negligible for all measured constituents. Correction factors for CSF creatine kinase and lactate dehydrogenase activities accounting for cellular contamination were developed. Total nucleated cell count was similar to counts in CSF of other species, but higher than values in healthy people. Differential leukocyte count in CSF was similar to that reported in CSF of other domestic animals: mostly lymphocytes, fewer monocytoid cells, and scant neutrophils. Cerebrospinal fluid protein concentration was higher than concentration reported for dogs, goats, and people, but was similar to values reported for horses. Beef cows had higher CSF total protein concentration than did dairy cows; also, beef cows had higher CSF gamma-globulin concentration. The concentration of sodium in CSF was slightly higher than the value in serum, and potassium concentration was lower than the value in serum. In contrast to studies of human beings, CSF osmolality was generally less than serum osmolality in the cows studied. Reference values for CSF electrolyte concentrations and osmolality are useful for diagnosis of salt poisoning and for assessment of the effects of fluid therapy. Magnesium concentration was lower in CSF, compared with serum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Detection of oligoclonal bands in cerebrospinal fluid from German Shepherd dogs with degenerative myelopathy by isoelectric focusing and immunofixation

BACKGROUND: Detection of intrathecal IgG synthesis is important in evaluating inflammatory diseases in the central nervous system. Isoelectric focusing (IEF) is currently the most sensitive method to demonstrate intrathecal IgG synthesis and may have diagnostic value for German Shepherd degenerative myelopathy (GSDM). OBJECTIVE: The objective of this study was to adapt a modified IEF and immunofixation method for the detection of oligoclonal bands in cerebrospinal fluid (CSF) from dogs with GSDM. METHODS: Serum and lumbar CSF samples were collected from 6 German Shepherd dogs clinically diagnosed with GSDM. Samples were also collected from 6 clinically healthy dogs for comparison. The concentration of IgG was measured by quantitative ELISA and the concentration of total protein was measured by the Bradford protein assay. The presence of oligoclonal bands was evaluated by use of modified IEF followed by immunofixation. RESULTS: The concentrations of total protein and IgG, and the IgG/total protein ratio in CSF samples, were not significantly different between GSDM patients and control dogs. Four GSDM patients had oligoclonal bands in their CSF based on IEF-immunofixation. No oligoclonal bands were found in CSF from control dogs. CONCLUSION: The results suggest that the detection of oligoclonal bands by IEF-immunofixation may have diagnostic value for GSDM, and support the idea that humoral immune responses may play a role in the pathogenesis of GSDM.     

广西部分地区猪群主要疫病免疫抗体监测和免疫效果分析

从广西5个地市33家养殖场随机采集免疫过猪繁殖与呼吸综合征(PRRS)、猪瘟(CSF)、猪伪狂犬病(PR)疫苗的血清样品393份,使用免疫胶体金方法对样品的抗体水平进行检测。结果显示,不同养殖规模的猪场抗体阳性率总体较高的是猪瘟,达75.06%;其次是猪伪狂犬病,为71.25%;而猪繁殖与呼吸综合征抗体最低,仅为42.24%。各种病原抗体在不同规模的猪场有较大的差别,规模猪场的猪群某些疫病的抗体未必比散养猪群高。通过对不同类型的疫苗所产生的抗体水平进行比较,结果发现,使用猪繁殖与呼吸综合征灭活苗和弱毒苗、猪瘟脾淋苗和细胞苗、猪伪狂犬病国产苗和进口苗免疫所产生的抗体水平没有明显差异。     

Virulence of recent and former classical swine fever virus isolates evaluated by their clinical and pathological signs

The clinical diagnosis of classical swine fever (CSF) still caused problems to the veterinarians during the last decade. The primary CSF outbreak was often detected too late and, meanwhile, the virus had spread. Consequently, the recent classical swine fever virus isolates (CSFV) were suspected to be of low virulence. The purpose of the study was to quantify the virulence of four recent CSFV by evaluating the clinical and pathological signs caused by different CSFV. Pigs of the same breed and age group were inoculated intranasally with CSFV from recent epidemics in European Union (EU) member states. The CSFV used are registered in the data base of the EU Reference Laboratory for CSF and belong to different genotypes: 2.1, 2.2 and 2.3 respectively. Clinical signs of CSF were evaluated by using a score system suggested previously (Mittelholzer et al., 2000: Vet. Microbiol. 74, 293). For the evaluation of pathological lesions, a new pathological score was introduced. The four CSFV tested here were classified as moderately virulent in general, although one CSFV may cause different clinical courses, ranging from highly virulent to avirulent. This indicates the importance of additional factors in the host animal for virulence. Differences in the clinical and pathological signs between these four recent CSFV were rather minor, emphasizing that the genetic typing of CSFV is absolutely essential. Differences towards former CSFV (e.g. reference virus strain Alfort 187) were more pronounced, especially regarding the onset and duration of the disease, the occurrence of skin haemorrhages and pathological lesions of kidney, subcutis and serosae. It is concluded that clinical diagnosis of CSF is rather difficult in pigs up to 14 days post-CSFV infection using these four CSFV, emphasizing the need for careful differential diagnosis and the laboratory investigation for CSF at an early stage.