美国农业部批准Synbiotics公司的Witness FeLV检测试剂盒

Synbiotics宣布美国农业部批准了它们关于Witness猫白血病病毒(FeLV)检测试剂盒的申请。这个试剂盒丰富了Synbiotics公司的Witness兽医诊断系列产品，这一系列有犬心丝虫病病原诊断试剂盒，猫犬怀孕诊断试剂盒。     

犬细小病毒套式PCR检测方法的建立及应用

为建立一种快速的犬细小病毒(CPV)病原检测方法,本试验根据GenBank上登录的CPV基因序列,在保守的VP2基因内部设计合成内外2对引物,建立了检测CPV的套式PCR方法。该方法对犬瘟热病毒(CDV)、犬冠状病毒(CCoV)、犬腺病毒(CAV)、犬副流感病毒(CPIV)的扩增结果均为阴性;该检测方法最低可以检测出0.1 ng的病毒核酸,第2次比第1次扩增的敏感性高100倍。建立的套式PCR方法具有良好的特异性、敏感性,可以准确快速检测出极低含量的CPV,将为CPV的病原检测及分子流行病学调查等提供一种高效、快速、特异、灵敏的检测方法。     

猫冠状病毒荧光RT-RAA检测方法的建立

根据猫冠状病毒(FCoV)7b基因序列,设计引物及探针,建立了FCoV荧光RT-RAA检测方法。该方法能够在39℃恒温条件下,20 min内对病原进行快速检测,与猫疱疹病毒、猫杯状病毒、猫细小病毒均无交叉反应;对病毒核酸的最低检测限为1.97×10~1 fg/μL。用套式PCR与建立的荧光RT-RAA对30份疑似FCoV感染临床样品进行检测,两种方法符合率为93.33%。结果表明,建立的FCoV荧光RT-RAA方法适用于FCoV快速检测。     

肉食动物细小病毒的检测方法

从猫、犬、貂、貉、兰狐和豹等多种肉食动物中分离到许多类似的细小病毒,并以其宿主分别称为:猫泛白细胞减少症病毒(FPLV)、犬细小病毒(CPV)、水貂肠炎病毒(MEV)、貉细小病毒(RPV)、兰狐细小病毒(BFPV)和豹细小病毒(LPV),这些细小病毒均具有细小病毒科及细小病毒属的典型特点,在形态、理化特性上极为相似,本文仅就上述几种细小病毒的检测方法做一简述,供科学研究、临床实践、海关检疫等有关人员参考借鉴。     

食源性人兽共患病病原活菌检测技术研究进展

食源性人兽共患病已成为威胁人类健康、食品安全和畜牧业发展的重要因素,因此发展快速、准确、高效的人兽共患病病原活菌检测技术非常必要。目前已有多种针对活菌的检测方法。本文比较分析了目前正在使用或开发的人兽共患病病原活菌检测技术。同时,利用PMA-q PCR检测技术,进行了副溶血性弧菌检测效果验证,认为该检测技术兼具检测速度快、灵敏度高、特异性强、成本低等优点,作为一种新型的活菌快速检测技术,具有广阔的应用前景。另外,一种新型的铂化合物检测技术也被证明可用于活菌检测。     

猪细菌性传染病病原检测新方法研究进展

细菌性传染病是生猪养殖业的严重威胁之一，近年来其发病情况呈现出新变化，对动物产品质量和公共卫生安全造成严重危害。对病原进行快速、准确的检测，是预防和控制猪病的有效手段，对养猪业发展和人畜健康具有重要意义。伴随生命科学技术的发展和学科间的交叉融合，传统的病原菌分离培养、普通PCR方法和早期免疫学等检测技术均朝着精确高效的方向不断发展革新，并以传统方法为基础建立了多种新型检测技术。文章重点介绍了猪细菌性传染病病原检测的几种新技术：以核酸分子生物学为基础的实时荧光定量PCR法（qPCR）、环介导等温扩增法（LAMP）、夹心DNA杂交（DNAH）等；以免疫学为基础的免疫胶体金标记技术、时间分辨荧光免疫分析法（TRFIA）、免疫磁珠分离法（IMS）等；以蛋白质分子生物学为基础的细菌质谱鉴定技术；以及适用于现场快速诊断的即时检测（POCT）技术，并分析了这几种新技术的优缺点，为不同条件的检测机构和研究部门在开展猪细菌性传染病检测和研究时选择相应的方法提供一定的参考，以期在猪病预防控制中发挥更加积极的作用。     

犬猫肠道菌群特点与营养调控研究进展

肠道菌群逐渐被认为是与犬猫健康密切相关的代谢活性器官，其参与营养物质的消化吸收、机体代谢及免疫等多种生理功能，决定着犬猫整体健康状态。食物营养素作为犬猫肠道菌群的生长基质，在决定肠道微生物组成及代谢方面起着重要作用。宠物食品中除碳水化合物、蛋白质和脂肪等营养成分外，也包括越来越多的微生物靶向成分，如益生菌和益生元等，这些营养素亦能调控犬猫肠道菌群结构及微生物代谢产物的生成，以达到改善宠物整体健康的目的。为此，本文就犬猫肠道菌群特点、功能以及饮食营养素对犬猫肠道菌群的影响进行综述，旨在为营养调控肠道菌群，改善犬猫健康及促进功能性宠物食品发展提供理论依据。 [关键词] 犬猫|肠道菌群|益生菌|益生元     

基于RPA-CRISPR/Cas12a技术快速检测犬猫皮肤癣菌方法的建立

旨在建立重组酶聚合酶扩增技术(recombinase polymerase amplification, RPA)结合CRISPR/Cas12a技术的荧光检测方法，以快速、灵敏、可视化检测犬猫皮肤癣菌。以犬小孢子菌、石膏样小孢子菌及须毛癣菌为研究对象，针对真菌内转录间隔区，设计并合成特异性RPA引物和CRISPR RNA(crRNA),建立可同时或分别检测上述皮肤癣菌的荧光检测方法，并评价其检测灵敏度，通过检测临床样本评价本检测方法的敏感性和特异性。结果表明：RPA-CRISPR/Cas12a在37℃、总反应时间30 min的条件下，对三种皮肤癣菌的检测灵敏度可低至单拷贝。对24份临床样本进行检测，以真菌培养和菌落测序结果作为标准，使用可同时识别犬小孢子菌、石膏样小孢子菌及须毛癣菌的皮肤癣菌crRNA(dermatophyte crRNA,crRNA-DM)和可特异性识别犬小孢子菌的犬小孢子菌crRNA(Microsporum canis crRNA,crRNA-Mc)参与RPA-CRISPR/Cas12a反应时，敏感性和特异性均为100%。RPA-CRISPR/Cas12a技术可同时...     

猫传染性腹膜炎病毒荧光定量RT-PCR检测方法的建立

为快速、灵敏、准确检测猫传染性腹膜炎病毒（feline infectious peritonitis virus,FIPV）,以FIPV N基因为靶序列设计合成特异性引物及TaqMan探针,建立了一种FIPV实时荧光定量RT-PCR检测方法,并对该方法的特异性、灵敏度和重复性进行了试验。结果表明：该方法灵敏度高,最低检测限为3.4 copies/μL;特异性强,与狂犬病毒、猫疱疹病毒、猫杯状病毒、猫细小病毒、犬流感病毒、犬瘟热病毒等均无交叉反应;重复性好,批内变异系数为0.99%~4.13%,批间变异系数为1.29%~4.64%。以上结果说明,本试验建立的实时荧光定量RT-PCR检测方法特异、敏感、重复性好,适用于FIPV的快速检测。     

多重核酸检测技术在动物疫病诊断和食品安全监测领域的研究进展

核酸检测技术是目前动物疫病诊断和食品安全监测的重要手段。多重核酸检测技术可同时检测多种病原,具有通量高、损耗低的优势,在疫病诊断和食品安全监测等领域的应用越来越广泛。目前已研发出多种多重核酸检测技术,包括聚合酶链式反应(polymerase chain reaction,PCR)多病原核酸检测技术、环介导等温扩增(loop-mediated isothermal amplification,LAMP)多病原核酸检测技术、生物芯片多病原核酸检测技术、高通量测序多病原核酸检测技术等。由于检测原理不同,不同方法之间的检测性能存在较大差异。本文通过概述基于PCR/RT-PCR、LAMP/RT-LAMP、生物芯片、高通量测序建立的多重核酸检测技术,全面分析其在动物疫病诊断和食品安全监测领域中的特点和具体应用策略,以期推进病原检测技术的创新和发展,为动物疫病诊断及食品安全监测提供技术支持。     

犬冠状病毒诊断技术研究进展

犬冠状病毒（canine coronavirus,CCoV）是导致犬胃肠道疾病的常见病原,与其他肠道病原共同感染时犬死亡率显著升高,严重影响养犬业的健康发展。快速、灵敏和特异的诊断技术对该病的防控起到至关重要的作用。除临床诊断外,常见的CCoV实验室诊断技术主要有病毒分离培养、电镜观察及血清学诊断技术,然而这些技术往往耗时长且工作量较大,分子诊断技术主要包括普通RT-PCR、实时荧光定量RT-PCR、纳米PCR等检测方法,这些诊断技术在准确性、敏感性及特异性上有极大提高,诊断效率也显著提高。近年来,随着分子免疫诊断技术与新型材料研发技术的迅速发展,新的检测方法应运而生,纳米金、核酸探针、芯片技术及纳米生物传感器等技术更加敏感、便捷、高效且都具有制备试剂盒的可能性,从而在诊断上具有简洁性和普适性。文章将针对CCoV的诊断技术进行全面综述,以期为CCoV诊断试剂的研发提供参考。     

新型分子诊断技术在动物疫病检测中的应用与发展

快速有效的病原体检测对于动物疫病的防控和治疗有着非常重要的作用。近年来基于PCR扩增技术的新型分子诊断技术与传统的分子诊断技术相比,对病原体的检测更加快速、特异、敏感、准确,逐步向着分子检测的自动化、高通量和现地使用方向发展。日渐成熟的新型分子诊断技术对于动物疫病检测与鉴别以及出入境检验检疫等领域有着广阔的应用前景,同时可进行大范围的流行病学调查和分析,为动物疫病的科学防控奠定基础。     

Current concepts in the diagnosis and treatment of brain tumours in dogs and cats

Intracranial tumours occur relatively frequently in dogs, and less commonly in cats. With the availability of computed tomography (CT) and magnetic resonance imaging, more accurate determination of the location and extent of brain tumours in companion animals has become possible. Following these advances in imaging, precise CT-guided stereotactic techniques for both tumour biopsy and intratumoral drug delivery have been developed for use in cats and dogs. Also, tumour identification methods, such as crush preparation examination, have facilitated rapid tumour identification. The use of improved diagnostic techniques has resulted in an increasing demand for effective therapies for brain tumours. While surgical removal and irradiation remain important treatment considerations in the management of brain tumours of cats and dogs, the development of gene therapy strategies for treatment of intracranial tumours offers much promise, although research in this area is still at an early stage.     

Evaluation of techniques for detection of small trematode eggs in faeces of domestic animals

This study was conducted to evaluate techniques for detection of small trematode eggs in faeces of dogs, cats and pigs. Faecal samples from dogs (n=80), cats (n=35) and pigs (n=114) were examined by Kato-Katz technique (KK), formalin-ether sedimentation technique (FE) and a method of combining: filtration, sedimentation and centrifugation, developed at DBL - Centre for Health Research and Development (former Danish Bilharziasis Laboratory) (DBL). Necropsy was performed on 38 dogs, 25 cats and 16 pigs and was considered as a gold standard method for evaluation of the techniques. The results showed 100% specificity for the three techniques. Lower sensitivity was seen for the KK-technique in dog samples in comparison to that for DBL- and FE-technique. The sensitivity of the three techniques was similar in cats and pigs. Based on these findings and practical issues, DBL-technique was chosen as most suitable because the eggs were easily detected and quantified. No toxic chemicals or special equipment were required in comparison with FE-technique that needs ether solution and thus fume cupboards which are often unavailable in local veterinary centres in Vietnam and other developing countries.     

Feline aelurostrongylosis and canine angiostrongylosis: a challenging diagnosis for two emerging verminous pneumonia infections

Aelurostrongylus abstrusus and Angiostrongylus vasorum (Nematoda, Metastrongyloidea) are causative agents of verminous feline and canine pneumonia. Both are presently emerging in several geographical areas and are of major clinical importance. Given the range of parasitic and non-parasitic pathogens which may cause cardio-respiratory distress in dogs and cats, the unequivocal specific diagnosis of both diseases is pivotal for unravelling their epidemiology and central to therapy and control. Several conventional clinico-pathologic, diagnostic imaging and parasitological approaches are currently used in practice, but all have major limitations in the specific diagnosis of these diseases. Serological methods have shown promise in the diagnosis of A. abstrusus and A. vasorum infections, but no clinically useful tests are currently available. Additionally, there have been major advances in the development of novel molecular diagnostic tools for feline aelurostrongylosis, but efforts to develop molecular diagnostic tests are still preliminary for angiostrongylosis. This article provides a review of A. abstrusus and A. vasorum infections in cats and dogs, focusing on the advantages and shortcomings of classical diagnostic methodologies and on present diagnostic advances as well as future perspectives instrumental to epidemiological and clinical studies.     

猫人工授精技术研究进展

人工授精技术是一种较先进的辅助生殖技术,可提高繁殖育种的效益,但因猫类较独特的生殖特点,其人工授精技术发展受限。近年来,科研人员和临床工作者借鉴其他物种相对成熟的人工授精技术,结合猫类生殖特点,对猫人工授精过程中精液采集、精液品质评估、精液保存、刺激母猫排卵方式及具体的人工输精等相关技术进行了优化和创新,如开发了新颖实用的尿导管法采精技术以采集猫精液;在传统精液品质评估的基础上,配合使用荧光染色技术、流式细胞技术及体外受精技术对猫精液品质进行评估;探索不同冷冻液配方及冷冻程序对猫精液保存效果的影响,并尝试使用超快速冷冻法保存公猫遗传物质;研究使用不同种类及不同剂量的激素对诱导母猫发情和排卵效果的差异,以及不同输精部位、方法及时机对母猫妊娠结果的影响等。尽管其中一些新技术与方法已在试验动物中彰显出了其实际应用潜力,但当下依旧还存在操作相对复杂、成本较高、效果低于预期等问题,导致目前猫人工授精技术的应用较局限,不如在犬或其他动物中应用广泛,未来还需更进一步研究和探索才能满足临床实际应用的需要。文章就上述猫人工授精技术中各个方面的研究进展进行了综述,以期为该技术的进一步研究和应用提供参考。     

Comparison of PCR, virus isolation, and indirect fluorescent antibody staining in the detection of naturally occurring feline herpesvirus infections.

Cats with clinical signs suggestive of ocular infection with feline herpesvirus type 1 (FHV 1) and cats without such signs were assayed by 3 methods to detect FHV. Comparison of polymerase chain reaction (PCR), virus isolation, and indirect fluorescent antibody staining techniques for the detection of FHV demonstrated higher sensitivity of PCR in detecting this common infectious agent of cats. Compared with PCR, sensitivity and specificity for virus isolation was 49% and 100%, respectively, and those of indirect immunofluorescence were 29% and 96%, respectively. FHV was detected in 13.7% of client-owned cats with conjunctivitis and in 31% of shelter cats with no ocular signs. The use of FHV PCR as a diagnostic test for FHV-associated disease is limited because of the occurrence of healthy carriers.     

Progress in development of immunocontraceptive vaccines for permanent non-surgical sterilization of cats and dogs

Each year, millions of cats and dogs are euthanized worldwide. There are insufficient resources to control shelter animals in developed countries, as well as feral cat and wild dog population levels, with current surgical sterilization techniques. Thus, population control of these animals will likely depend on the development of new non-surgical methods for cat and dog sterilization. One promising area of research is the development of contraceptive vaccines, or immunocontraceptives. In this article, previous approaches aimed at developing contraceptive vaccines will be reviewed, with a focus on those most related to sterilization of cats and dogs. There are a number of steps in reproduction that have been, or could be, targeted by the immune system, and the advantages and obstacles for inducing immunity to each of these will be discussed. Our current understanding of how these vaccines cause sterility, and our current ability to dissect these mechanisms in cats and dogs, also will be discussed.     

Nasal hydropulsion: a novel tumor biopsy technique

Intranasal tumors of dogs and cats pose a diagnostic and therapeutic challenge for small animal practitioners. Multiple nasal biopsy techniques have been described in the past. This report describes a simplified flushing technique to biopsy and debulk nasal tumors, which often also results in immediate clinical relief for the patient. Based on the results of this retrospective study, the authors recommend high-pressure saline hydropulsion as a minimally invasive diagnostic, and potentially therapeutic, technique for nasal tumors in dogs and cats.     

Campylobacteriosis in dogs and cats: a review

Campylobacter species are commonly isolated from faecal samples collected from dogs and cats, with the most prevalent species being C. upsaliensis, C. helveticus, and C. jejuni. Although the majority of dogs and cats are subclinically infected, some will develop mild to moderate enteritis. Immature animals, animals from intensive housing backgrounds, and animals with concurrent disease are especially predisposed to infection and the development of clinical signs. Bacterial culture methods applied in diagnostic laboratories remain biased to C. jejuni and C. coli detection, but molecular methods to diagnose Campylobacter spp. infections in dogs and cats have become widely available and can aid rapid and accurate diagnosis. Multilocus sequence typing has also been developed for subtyping different strains and has been used in epidemiological investigations. In the majority of cases, clinical signs are self-limiting and antimicrobial treatment is not warranted. Campylobacter spp. isolated from dogs and cats have shown resistance to commonly used antimicrobials, so antimicrobial therapy should only be administered where this is justified. Contact with dogs and cats is a recognised risk factor for human campylobacteriosis, thus people living or working in close contact with cats and dogs should be made aware of the zoonotic organisms these animals can shed.