Rehabilitation for the neurologic patient.

A properly designed rehabilitation program should be an important component of the treatment plan of animals with neurologic disease.Such a program should be designed in conjunction with appropriate treatment of the underlying problem and after special consideration of the origin of the neurologic problem, the severity of the signs, the cause of the signs, their anticipated progression, and the needs of the owner and the pet. This article describes the pathophysiology of injury and recovery in the central and peripheral nervous systems, assessment of the neurologic patient, data on the prognosis and expected course of recovery for a variety of different diseases, and rehabilitation exercises appropriate for neurologic patients.     

Update on molecular techniques for diagnostic testing of infectious disease.

The era of diagnostic molecular biology has arrived for small animal clinicians, and it is a near certainty that assays such as the PCR and RT-PCR will become more widely available for a wider array of infectious agents. Already there is an extensive list of infectious diseases of dogs and cats that have been investigated with molecular tools. A partial list is included in box 1. An understanding of the advantages and disadvantages of the molecular techniques and some of the questions these techniques can answer for clinicians can serve practitioners well in their approach to the diagnosis of infectious diseases in dogs and cats. It is likely that additional applications of these tools to small animal medicine will become apparent as investigators use and refine them for their research purposes, or as new uses emerge from human medical applications. Clinicians also are likely to reap the benefits of this knowledge. Because samples often are acquired easily from clinical patients in most practice settings, access to these tools puts all clinicians in the group of discoverers of new, or variations of, infectious diseases and their clinical manifestations.     

Bayes and diagnostic testing

Interpretation of the result of a diagnostic test depends not only on the actual test result(s) but also on information external to this result, namely the test's sensitivity and specificity. This external information (also called prior information) must be combined with the data to yield the so-called updated, posterior estimates of the true prevalence and the test characteristics. The Bayesian approach offers a natural, intuitive framework in which to carry out this estimation process. The influence of the prior information on the final result may not be ignored. Guidance for the choice of prior information not in conflict with the data can be obtained from a set of statistics and indices (DIC, p(D), Bayes-p).     

The diagnostic and therapeutic approach to the patient in acute congestive heart failure

The patient presented in acute congestive heart failure represents a diagnostic challenge. Life-threatening compromise of respiratory or cardiovascular function renders the patient vulnerable to the stresses associated with many diagnostic tests. Before risking any test in this situation, it is important to understand the information that each test will give, and how that will impact the diagnostic and therapeutic approach in this crucial early phase of patient management. This article describes the diagnostic tests commonly used in the evaluation of patients in acute heart failure and the information that each provides, and discusses how this information can be used to guide therapy. Pericardiocentesis is discussed in detail.     

奶牛附红细胞体的分类鉴定及诊断方法的建立

为了从分子水平上确定奶牛附红细胞体（E．wenyoni）的分类学地位及建立奶牛附红细胞体感染诊断方法。本研究通过利用原核生物16S rRNA基因通用引物，对分离得到的奶牛附红细胞体（广西株，E．wenyoni-GX）进行16SrRNA基因的克隆及测序。并建立系统发育进化树；同时根据测序结果设计诊断引物，建立奶牛附红细胞体感染的PCR诊断方法。结果扩增出长约1．5kbp的奶牛附红细胞体的16SrRNA基因片段；特异性试验和敏感性试验表明。所建立的PCR方法与常见支原体、细菌及原虫元交叉反应，能检测奶牛附红细胞体最低DNA量为0．145fg。通过试验结果分析，建议将奶牛附红细胞体这类血营养菌划归入支原体科、支原体属；同时，所建立的PCR诊断方法是特异、敏感、快速的，可应用于临床检测。     

饲料中牛羊源性成分PCR方法定性检测能力验证计划的实施和结果研究

"饲料中牛羊源性成分PCR方法定性检测"是由国家认证认可监督管理委员会(CNCA)组织的A类能力验证计划.通过对本次能力验证计划的实施和对结果的分析研究,了解了国内饲料中牛羊源性成分PCR检测领域的整体水平以及各实验室在检测中存在的差异.     

猪瘟、猪圆环病毒病和猪繁殖与呼吸综合征寡核苷酸芯片诊断方法的建立与初步应用

猪繁殖与呼吸综合征病毒（PRRSV）、猪瘟病毒（CSFV）和猪圆环病毒Ⅱ型（PCV-2）是引发猪繁殖障碍的主要病原,建立其基因芯片快速检测体系,对开发猪繁殖障碍快速检测试剂盒具有重要意义。根据GenBank中已发表的PRRSV、CSFV和PCV-2的病毒基因组序列,设计合成特异性引物和特异性较强的60mer的寡核苷酸探针,并将探针按所设计阵列固定于表面经氨基化修饰的玻片上,制备出寡核苷酸芯片。通过引物标记及特异性验证,建立了带标记引物的多重PCR检测体系,从而产生大量可与寡核苷酸探针特异性互补的带标记的DNA片段。将标有荧光染料的扩增产物与芯片上寡核苷酸探针杂交,扫描、分析芯片上荧光信号。结果表明,芯片上各样本对应探针位点呈现阳性荧光信号,而阴性对照和空白对照则基本不能检测到荧光信号。分别用基因芯片检测方法和PCR/RT-PCR检测60份临床病料,两者符合率高达92%,表明该检测技术能够用于临床病料的检测及快速诊断PRRSV、CSFV和PCV-2。     

PCR as a diagnostic tool for brucellosis

Numerous PCR-based assays have been developed for the identification of Brucella to improve diagnostic capabilities. Collectively, the repertoire of assays addresses several aspects of the diagnostic process. For some purposes, the simple identification of Brucella is adequate (e.g. diagnosis of human brucellosis or contamination of food products). In these cases, a genus-specific PCR assay is sufficient. Genus-specific assays tend to be simple, robust, and somewhat permissive of environmental influences. The main genetic targets utilized for these applications are the Brucella BCSP31 gene and the 16S–23S rRNA operon.

Other instances require identification of the Brucella species involved. For example, most government-sponsored brucellosis eradication programs include regulations that stipulate a species-specific response. For epidemiological trace back, strain-specific identification is helpful. Typically, differential PCR-based assays tend to be more complex and consequently more difficult to perform. Several strategies have been explored to differentiate among Brucella species and strains, including locus specific multiplexing (e.g. AMOS-PCR based on IS711), PCR-RFLP (e.g. the omp2 locus), arbitrary-primed PCR, and ERIC-PCR to name a few. This paper reviews some of the major advancements in molecular diagnostics for Brucella including the development of procedures designed for the direct analysis of a variety of clinical samples. While the progress to date is impressive, there is still room for improvement.     

常规PCR和巢式PCR法鉴定牛早期胚胎性别体系的建立和优化

试验利用牛Y染色体重复序列作为雄性特异性引物,_以肿蔼坏死因子(TNFα)为内标引物建立多重PCR和多重巢式PCR体系,进行牛早期胚胎性别鉴定.共设计4对引物-Y染色体重复序列外引物和内引物,其扩增片段大小分别为534 bp和480bp,肿瘤坏死因子外引物和内引物,扩增片段大小分别为357 bp和272 bp.结果表明,4对引物均有很高的特异性和稳定性;多重PCR体系灵敏度为50 pg(约8个细胞),多重巢式PCR体系灵敏度为10 pg(约2个细胞),故多重巢式PCR体系更适合于牛胚胎性别鉴定.     

鸭病毒性肝炎病毒、禽流感病毒和新城疫病毒三重PCR诊断方法的建立及应用

根据GenBank中已登录的鸭病毒性肝炎病毒（DHV）、禽流感病毒（AIV）和新城疫病毒（NDV）的基因序列,设计了3对特异性引物,建立了DHV、AIV和NDV的多重PCR鉴别诊断技术,并对体系进行了优化和特异性、敏感性试验。结果表明,该体系所扩增的3种病毒基因片段大小分别为174bp（DHV）、264bp（AIV）和362bp（NDV）,且特异性、敏感性良好,能够分别检出1pg AIV、10pg DHV和10pg NDV的病毒RNA。对鸭临床样品的检测结果表明,该方法与病毒分离鉴定符合率90%以上,可用于临床样品的快速检测。     

Systemic neosporosis in a dog treated for immune‐mediated thrombocytopenia and hemolytic anemia

A 4‐year‐old male Toy Poodle was presented to the Small Animal Veterinary Hospital of the Faculty of Veterinary Medicine of the Autonomous University of Mexico (FMVZ, UNAM) because of depression, lethargy, and hemorrhages involving several areas of the skin and around the eyes. Hematology data and a bone marrow analysis suggested hemolytic anemia and immune‐mediated thrombocytopenia. The dog was treated with prednisone, and after one month the hematology variables improved. However, the dog's clinical condition inexplicably worsened and it was euthanized. On necropsy, there were no relevant findings. However, in histology, multifocal lymphoplasmacytic and histiocytic meningoencephalitis and necrosis, and a protozoan cyst in the cerebellum were identified. In addition, moderate multifocal lymphoplasmacytic and necrotizing pancreatitis, hepatitis, myocarditis, and diffuse lymphoplasmacytic enteritis were observed. Immunohistochemistry of the cerebellum, liver, pancreas, and intestine with a specific antibody against Neospora caninum confirmed the diagnosis of systemic neosporosis. The systemic neosporosis in this dog was most likely caused by reactivation of latent parasites due to prednisone administration during the one month of treatment. It should be kept in mind that in dogs being treated with immunosuppressants for immune‐mediated conditions, opportunistic parasites, such as Toxoplasma gondii and N caninum, can be reactivated from a latent state, as it probably happened in the present case.     

Repeat patient testing shows promise as a quality control method for veterinary hematology testing

Background

Repeat patient testing‐based quality control (RPT‐QC) is a potential method for veterinary laboratories (eg, that have a limited budget for quality commercial control material [QCM] or that wish to use material with a species‐specific matrix).

Objectives

To determine whether total error (TE_a), probability of error detection (Ped), and probability of false rejection (Pfr) similar to that achievable with QC materials can be controlled using RPT‐QC

Methods

Control limits (WBC, RBC, HGB, HCT, MCV, and PLT) for the Advia 120 (n = 23) and scil Vet ABC (n = 22) were calculated using data from normal canine specimens from a routine caseload. Specimens were measured at accession and again after 24 hours. Control limits were validated using 23 additional canine specimens tested similarly. Achievable TEa, Ped, and Pfr were investigated using the Westgard EZRules3 and compared to those achievable with commercial QCM.

Results

Theoretical performance of RPT‐QC and commercial QCM‐QC are similar for 1‐3s with both n = 1 and 1‐3s with n = 2 for all measurands and both instruments. Achievable TE_a values for RPT‐QC were close to ASVCP recommendations for most measurands; exceptions were PLT (both instruments) and WBC (scil Vet ABC).

Conclusions

Repeat patient testing‐based quality control advantages include a species‐specific matrix, low‐cost, and absence of QC material deterioration over time (since a fresh specimen is used each day). A potential disadvantage is daily access to normal canine specimens. A challenge is determining control limits, which has a subjective element. Further study is needed to confirm actual RPT‐QC performance and to determine if RPT‐QC with abnormal patient specimens is feasible.     

Combining anti‐IgM and IgG immunoassays for comprehensive chikungunya virus diagnostic testing

Chikungunya virus (CHIKV) is a mosquito‐borne pathogen that causes CHIKV fever. Definitive diagnosis is crucial for patients experiencing symptoms similar to other arboviral diseases because they can vary in clinical consequences. An increasing number of patients experience long‐term rheumatic effects of CHIKV infection, but these cases may not be optimally detected by molecular assays and anti‐CHIKV IgM ELISAs (M‐ELISAs) used for confirmation and screening, respectively. The subsequent confirmatory serological test, the plaque reduction neutralization test (PRNT), is laborious and time‐consuming. In this study, we evaluated a new diagnostic algorithm in which the M‐ELISA is conducted in parallel with an anti‐CHIKV IgG ELISA (G‐ELISA) and observed that the Euroimmun M‐ELISA combined with the Euroimmun G‐ELISA or the Abcam G‐ELISA exhibited excellent sensitivity and specificity for CHIKV. The combinations demonstrated perfect and near perfect inter‐rater agreement with the PRNT, respectively, suggesting their potential to be used as alternatives to the confirmatory serological PRNT assay for CHIKV.     

Consensus recommendations on diagnostic testing for the detection of paratuberculosis in cattle in the United States

The report provided here contains a simplified set of diagnostic testing recommendations. These recommendations were developed on the basis of research funded by the USDA-Animal and Plant Health Inspection Service-Veterinary Services through a cooperative agreement. The report is intended to provide simple, practical, cost-effective consensus testing recommendations for cattle herds that are not enrolled in the US Test-Negative Program. The information has been reviewed by paratuberculosis (Johne's disease) experts at the USDA and academic centers as well as stakeholders in various segments of the cattle industry. The recommendations were accepted by the National Johne's Working Group and Johne's Disease Committee of the US Animal Health Association during their annual meetings in October 2006. The report is intended to aid veterinarians who work with cattle producers in the United States. The recommendations are based on information available up to October 2006. There is a paucity of large-scale, high-quality studies of multiple tests conducted on samples obtained from the same cattle. It is understood that there may be special circumstances that require deviation from these recommendations. Furthermore, as new information becomes available and assays are improved and their accuracy is critically evaluated, changes to these recommendations may be necessary.     

Application of the polymerase chain reaction (PCR) in veterinary diagnostic virology

The polymerase chain reaction has become an important diagnostic tool for the veterinary virologist. Conventional methods for detecting viral diseases can be laborious or ineffective. In many cases PCR can provide a rapid and accurate test. In this article we explain the basic principles of PCR and supply a reference list of its uses in diagnostic veterinary virology.Abbreviations BLV bovine leukaemia virus - BVDV bovine viral diarrhoea virus - DNA deoxyribonucleic acid - ds double-stranded - ELISA enzyme-linked immunosorbent assay - PCR polymerase chain reaction - RNA ribonucleic acid - ss single-stranded - TK thymidine kinase     

A longitudinal study to evaluate the diagnostic potential of a direct faecal quantitative PCR test for Johne's disease in sheep

A longitudinal study was carried out to evaluate the diagnostic potential of the previously developed direct faecal real-time quantitative PCR (QPCR) assay (Kawaji et al., 2007) for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) infected sheep. Of the 58 sheep, 38 were orally inoculated with MAP, while 20 controls were maintained separately from the infected group throughout the trial. All animals were tested by QPCR, faecal culture and serum ELISA pre-inoculation and at 4, 8 and 13 months post-inoculation, and were necropsied at 13 months post-inoculation. Eighteen out of 38 inoculated sheep were detected by QPCR to be shedding MAP in faeces at 4 months post-inoculation, while only one sheep was positive in faecal culture at this time point. At 8 months post-inoculation, MAP DNA was detected in faeces of all inoculated sheep by QPCR, while MAP organisms were isolated from only 34% of the inoculated animals by faecal culture. The QPCR results for faecal samples that were collected at necropsy demonstrated that faecal QPCR was more sensitive than culture of intestinal tissues for MAP. The QPCR assay was confirmed to be a sensitive and specific ante-mortem diagnostic test for MAP in sheep, circumventing faecal culture which is a less sensitive and highly time consuming test. Quantification of MAP DNA in faeces by QPCR may provide immediate information to estimate the stage of the infection as well as the risk of transmission from infected animals.     

人参皂苷及其衍生物体内抗马立克氏病毒的间接免疫荧光观察与PCR检测

为了探讨人参皂苷及其衍生物体内抗马立克氏病毒的作用机理.用马立克氏病毒人工感染雏鸡模型,采取人参皂苷及其衍生物口服,通过间接免疫荧光试验动态观察人参皂苷及其衍生物能否减少MDV抗原在组织中的分布;通过PCR检测看人参皂苷及其衍生物能否减少MDV的出现.结果显示:人参皂苷组、衍生物组被检组织的阳性细胞数量明显比盐酸吗啉胍阳性对照组被检组织的阳性细胞数量少;MDV病毒核酸的PCR检测显示,药物没有阻止病毒对组织的感染.结果表明,人参皂苷及其衍生物抗马立克氏病毒效果要优于盐酸吗啉胍.