间接荧光抗体法快速检测病禽组织中的新城疫抗原

利用间接荧光抗体法(IFA)对用新城疫病毒(NDV)攻毒鸡胚的卵黄、胚体、尿囊液、尿囊膜和人工感染鸡、自然病死鸡的心、肝、脾、肺、肾、脑、盲肠扁桃体、法氏囊等8种脏器进行检测。结果鸡胚收集液、6例人工感染和15例自然病例的脏器检测结果均为阳性;脏器以肝、脾、肺、脑的检出率最高,检出效果最好。     

鸡败血霉形体抗原基因的筛选和DNA电泳鉴定

应用鸡败血霉形体抗血清筛选鸡败血霉形体Ｓ６株基因文库,得到１７９个阳性克隆,不足重组噬菌体的１％。反复筛选后,获得５个强阳性克隆。抽提ＤＮＡ酶切鉴定,发现２个克隆的外源片段在５ｋｂ左右,另２个在２．５ｋｂ左右,一个为３．６ｋｂ,为特异抗原的制备和基因工程疫苗的研究打下良好基础。     

蛋白质免疫印迹快速测定猪肺炎支原体抗原含量

为建立快速测定猪肺炎支原体抗原含量的方法,试验应用P46单抗建立不同CCU含量的猪肺炎支原体抗原蛋白免疫印迹图谱,测定不同批次的猪肺炎支原体抗原CCU含量,并与Western blot(WB)测定的CCU进行比较。结果显示:曝光5 s,107 CCU/mL含量以上的猪肺炎支原体抗原在46 kDa处有条带,且条带粗细、明暗程度随着CCU含量的增高而增强;中试生产的7批抗原中,有3批抗原CCU和WB 2种方法测定的含量相同,两批抗原测定的含量基本相同;两批抗原测定的含量有差异,但差值在1.5~5.0倍之间。结果表明,WB可用于猪肺炎支原体抗原含量的快速测定。     

用完整菌体作抗原包被进行ELISA检测激光照射前后血清抗体含量的初步研究

本研究用改良ELISA法检测激光照射前后抗大肠杆菌血清抗体的含量。结果表明:利用完整菌体作抗原包被进行ELISA试验,完全符合一般可溶性抗原的ELISA规律;并与传统血清学方法凝集反应进行了比较。激光照射穴位后使血清中抗体的含量增加,提示激光照射后能增强机体的免疫功能。     

牛分技杆菌胞浆特异蛋白质抗原的制备和检测

将牛分枝杆菌Mycobacterium boyis S_(10)株培养于苏通培养基,经低温高速离心,超声波裂解,Sephadex G200柱层析,获得3个峰,制备到3种胞浆蛋白质抗原,经同奶牛结核病分枝杆菌阳性血清。阴性血清及各种分枝杆菌高免牛血清的ELISA检测,并与牛PPD,聚合OT等抗原做了比较,以第二峰的胞浆蛋白质抗原的特异性为最好。     

Distinct antigen recognition pattern during zoonotic visceral leishmaniasis in humans and dogs

Leishmania infantum is a causative agent of endemic zoonotic visceral leishmaniasis (VL) in regions of South America and the Mediterranean. Dogs are the major reservoirs for L. infantum in these regions, and control of disease in dogs could have a significant impact on human disease. Although dogs share many symptoms of VL with humans as a result of L. infantum infection, they also show some unique clinical manifestations, which are often a combination of visceral and cutaneous leishmaniasis, suggesting different mechanisms of disease development in dogs and humans. Here, we compare antibody responses of dogs and humans with VL to various defined leishmanial antigens. Parasite lysate and K39, the two most commonly used antigens for serodiagnosis of VL, detected the highest levels of antibodies in both humans and dogs with VL, whereas the recognition patterns of these antigens were distinct between the hosts. Among other defined antigens tested, LmSTI1 and CPB detected higher levels of antibodies in dogs and humans, respectively. These results indicate there is a difference between humans and dogs in antigen recognition patterns during VL. We infer that different strategies may need to be used in development of vaccines and diagnostics for humans and for dogs. In addition, we show a correlation between antibody titers to several antigens and severity of clinical symptoms during canine VL.     

Optimizing methods in immunocytochemistry: one laboratory's experience

Abstract: The addition of immunocytochemical staining procedures to a diagnostic cytology service enables greater specificity of interpretation for many common disease conditions, especially neoplastic diseases. However, well‐tested immunohistochemical techniques may require modification for cytologic specimens, and other considerations are necessary when working with air‐dried cells. In this article, we describe our experience in evaluating options for sample transport and handling, and discuss methods for obtaining control cells from a variety of tissues for use in immunocytochemical staining. Important immunocytochemical principles and techniques, including fixation, antigen retrieval, and use of primary and secondary antibodies in manual and automated staining systems are described as used in our laboratory for cytologic specimens. Although we emphasize methods relevant to diagnostic laboratories receiving samples from external clients, the information is also applicable to any laboratory interested in adding or enhancing immunocytochemical services.     

鸡结核的病理形态学和抗原定位的观察

本文报道了９８只海塞克商品蛋鸡，经禽结核菌素变态反应试验，细菌染片检查、分离培养、生化鉴定、诊断为禽型结核。对这９８只鸡全部进行了尸体剖检，取１８只做了组织学观察，证明鸡结核病灶出现最多的器官是肝（９２％）、脾（７３％）、肠（７３％）；鸡结核病很少形成钙化灶。     

不同方法对鸡胚和法氏囊中囊病病毒抗原含量的检测比较

用单抗介导的抗原捕获－酶联免疫吸附试验（AC－ELISA），琼脂扩散试验（ACP）快速诊断试纸条（ISK）对感染同株强毒或弱毒囊病病毒（IBDV）的鸡胚和鸡法氏囊中的病毒抗原效价进行了检测比较，结果表明，上述3种方法测得法氏囊中温毒IBDV抗原效价分别为10^6.0(AC-ELISA),2^-3.0(AGP)t 10^-3.0，测得鸡胚中强毒IBDV抗原效价依次为10^-1.0,0和0。强毒株接种鸡胚后，只有第1代鸡胚组织可被AC－ELISA测到低效价的IBDV抗原，而第2代以后则未能测到病毒抗原，弱毒疫苗株IBDV感染鸡后，法氏囊中可测到低效价抗原，但感染鸡胚后则无可测性病毒抗原。     

Ag85B lentivirus inhibits expression of TSLP and TSLP receptor in human bronchial epithelial cells

AIM: To investigate the effect of Mycobacterium tubeculosisantigen （Ag85B） lentivirus on the expression of thymic stromal lymphopoietin (TSLP) and TSLP receptor (TSLPR) in cultured normal human bronchial epithelial cells (NHBEC) in vitro. METHODS: Ag85Bgene was subcloned into 293T cells with recombinant plasmid to yield the Ag85B lentivirus. The NHBEC were cultured and transfected with Ag85B lentivirus, NS control and empty lentivirus control using Lipofectamine 2000. The expression of Ag85B, TSLP and TSLPR in NHBEC at mRNA and protein levels was confirmed by real-time PCR and Western blotting.RESULTS: Ag85B lentivirus was constructed with a viral titer of 2×1011 TU/L and the Ag85B expression was detected. As compared with control group and empty lentivirus group, the mRNA expression of TSLP in Ag85B lentivirus group was found to slightly decreased, and the mRNA expression of TSLPR and the protein expression of TSLP and TSLPR were significantly decreased in Ag85B lentivirus group (P<0.05). CONCLUSION: The constructed Ag85B lentivirus antagonizes the expression of TSLP and TSLPR in cultured NHBEC, suggesting that Ag85B inhibits the initiation of asthmatic inflammation by blockage of TSLP-TSLPR pathway.