酶标单抗阻断ELISA检测鸡白痢和鸡伤寒抗体

以辣根过氧化物酶标记的沙门氏菌O9单抗3-47-0与包被的鸡白痢沙门氏菌脂多糖抗原建立了一种抗体阻断EL ISA以检测鸡白痢和鸡伤寒沙门氏菌感染。该方法基于待检血清样品抑制酶标单抗与脂多糖的结合,以检测鸡白痢和鸡伤寒特异性抗体。对72份已知鸡白痢阳性血清(抑制率为(98.5±4.0)%)、54份已知鸡白痢阴性血清(抑制率为(15.3±8.0)%)、42份SPF鸡血清(抑制率为(0.8±7.2)%)检测的结果表明,该方法具有很强的区分能力。在人工感染试验中,从第2周开始,该方法能从全部鸡只中检测到特异性抗体,比全血玻板凝集试验(PAT)早1周时间。对344份种鸡血清的检测结果表明,单抗阻断EL ISA比PAT特异性好、敏感性高。另外,该方法也能检测出与鸡白痢和鸡伤寒沙门氏菌有共同抗原的沙门氏菌感染产生的抗体。     

抗鸡白痢沙门氏菌单抗及其在鸡白痢病检疫中的应用研究

本研究应用淋巴细胞杂交瘤技术制备了一组针对鸡白痢沙门氏菌O_9抗原的单抗。以单抗3-47-26酶结合物和鸡白痢沙门氏菌脂多糖抗原建立了一种抗体竞争ELISA试验以检疫鸡白痢病,该法通过样品抑制酶标单抗与LPS的结合,以检测鸡白痢抗体。试验结果表明,79份鸡白痢阳性血清具有显著抑制作用(95.0±4.3％),而130份鸡白痢阴性血清以及48份SPF鸡血清抑制不明显,它们的抑制率分别为12.3±10.6％和6.2±5.5％。对94份种鸡血清的检疫结果表明,抗体竞争ELISA比PAT、AGPT、PHA敏感和特异,并进一步讨论了该法在血清流行病学调查方面的应用前景。     

鸡白痢诊断抗原的研制

应用标准型C79-1和C79-7鸡白痢鸡伤寒沙门氏菌标准株,制备了鸡白痢鸡伤寒平板凝集试验染色诊断抗原。该抗原特异性强,与鸡新城疫、鸡传染性鼻炎、鸡支原体等9种鸡血清无交叉反应,敏感性高,稳定性好,4℃保存36个月,检测效果不变,应用此抗原检测鸡白痢鸡伤寒沙门氏菌抗体,检测快速,操作简便,适于大规模临床样品检测。     

单抗竞争EILSA试验及其在鸡白痢病检疫中的应用

应用针对沙门氏菌 O_9抗原的单抗3-47-26酶结合物和鸡白痢沙门氏菌脂多糖抗原建立了一种单抗竞争 ELISA试验,以检测鸡白痢抗体.试验结果表明,79份鸡白痢阳性血清具有显著抑制作用,而130份鸡白痢阴性血清以及48份 SPF 鸡血清抑制不明显.对94份种鸡血清的检疫结果表明,单抗竞争 ELISA 比 PAT、AGPD、PHA 特异性好、敏感性高,文中进一步讨论了该法的应用前景.     

基于pegB基因的鸡白痢沙门氏菌、鸡伤寒沙门氏菌分子鉴别方法的建立

为建立一种能快速、准确、简便区分鸡白痢沙门氏菌和鸡伤寒沙门氏菌的方法，研究通过对鸡白痢沙门氏菌和鸡伤寒沙门氏菌pegB序列进行分析，发现存在3个单核苷酸多态性位点，基于此建立了一种特异性PCR方法用于鉴别鸡白痢沙门氏菌和鸡伤寒沙门氏菌。结果显示，该方法特异性强、耗时短，检测限为10.14 pg/反应。临床样品检测表明，该方法不仅具有良好的特异性，且操作比传统方法更为简便。该方法的建立为准确区分鸡白痢沙门氏菌和鸡伤寒沙门氏菌提供了实用技术，将在分子检测中发挥重要作用。     

鸡白痢鸡伤寒多价染色平板凝集抗原的研制与应用

应用标准型C79-1和C79-7鸡白痢鸡伤寒沙门氏菌标准株,制备了鸡白痢鸡伤寒平板凝集试验染色诊断抗原。该抗原特异性强,与鸡新城疫、鸡传染性鼻炎及鸡支原体等9种鸡血清无交叉反应,敏感性高,稳定性好,4℃保存36个月,检测效果不变,应用此抗原进行平板凝集反应检测鸡白痢鸡伤寒沙门氏菌抗体,操作简便,检测快速,适于普及。     

减毒鸡沙门氏菌疫苗株97A安全性评价

鸡白痢（pullorum disease）和禽伤寒（typhus avium）是分别由鸡白痢沙门氏菌与鸡伤寒沙门氏菌感染引起鸡的一种败血性疾病，我国对其控制主要采用检疫淘汰的方法，并使用抗生素预防和治疗。但就目前现状而言，采取淘汰清除措施来消灭沙门氏菌，条件还不成熟，加之抗生素治疗也存在诸多弊端，因此，有必要探寻新的防制途径。     

2006--2012年广西鸡沙门氏菌感染情况调查

运用分子生物学、血清学以及病原分离鉴定等多种方法，对2006～2012年广西规模养殖场、农村散养鸡以及活禽市场鸡血清和组织样本进行鸡白痢-伤寒沙门氏菌抗体和沙门氏菌病原检测和鉴定。结果发现，血清样本中规模场鸡白痢伤寒抗体阳性率较低，平均为2．16％，其中种公鸡的阳性率平均为1．55％，核心群母鸡阳性率平均为1．66％，生产群母鸡阳性率平均为4．16％；但广西农村散养鸡和活禽市场鸡白痢-伤寒沙门氏菌的污染情况不容乐观，其中农村散养鸡白痢-伤寒沙门氏菌阳性率为21．78％～24％，平均为22．52％（320/1421），活禽市场鸡阳性率为20．81％～33．2％，平均达到27．51％（2800/10177）；发病鸡的组织样品沙门氏菌平均阳性率为4．9％（29/592），阳性株以伤寒沙门氏菌为主，共13株占44．83％（13/29），其次为亚利桑那沙门氏菌8株，占27．59％（8/29），第三种是沙门氏菌的某些亚种（无法定型），最少为猪霍乱沙门氏菌2株，占6．9％（2/29）。     

鸡白痢沙门氏菌感染情况的调查

鸡白痢是由鸡白痢沙门氏菌引起的一种败血性细菌性疾病 ,它能垂直传递和水平传播。为了控制本病 ,对种鸡群进行净化是最根本的措施。鸡白痢净化的主要方法是检测淘汰阳性种鸡和防止再次感染。检测方法主要是采用现场全血平板凝集试验。为了最大限度检出和淘汰阳性鸡 ,连续进行了 4次检测。1　材料和方法鸡白痢平板凝集抗原 :由中国兽药检察所生产的鸡白痢鸡伤寒多价染色抗原。鸡白痢阳性和阴性对照血清 :由中国兽药检察所提供。检测方法和判断标准 :全血平板凝集试验。检测时机 :1 6周龄开始 ,全群普检 ,淘汰阳性鸡 ,如阳性率大于 0 .2 % ,…     

应用单抗协同凝集试验检疫鸡白痢

鸡白痢是经卵传播的疫病,检疫净化病原是普遍采用的方法。目前世界各国多采用鸡白痢鸡伤寒多价染色抗原全血平板快速凝集反应。凝集抗体的产生,通常在感染之后3～10天,最早3～4周。4周龄以下的雏     

后备种鸡血清中沙门氏菌病抗体的检测

采用全血平板凝集试验,对重庆市某种鸡场1113 只3月龄后备种鸡进行鸡白痢抗体检测，结果：阳性率为10.3%，其中X品系种公鸡阳性率为3.6%、种母鸡阳性率为1.3%，Y品系种公鸡阳性率为17.6%、种母鸡阳性率为11.8%。试验说明该鸡场后备种鸡中沙门氏菌病感染严重；不同品系、不同性别后备种鸡感染程度差异较大。     

应用斑点免疫金渗滤法诊断犬弓首蛔虫病

根据免疫渗滤原理,采用犬弓首蛔虫抗原蛋白和胶体金标记SPA,建立抗犬弓首蛔虫抗体的斑点免疫金渗滤法(D IGFA)检测200份犬血清样本。结果:阳性检出率为8.00%(16/200);血清样本对应犬粪便用饱和盐水漂浮法检查虫卵,阳性检出率为2.0%(4/200)。结论:斑点免疫金渗滤法具有操作简便、灵敏度高、特异性强的优点,可用于犬弓首蛔虫病的早期、快速检测。     

鸡源大肠杆菌和沙门氏菌的分离鉴定及耐药分析

为探究甘肃省民勤县某饲养场肉仔鸡腹泻的病原及复方中草药制剂和抗生素分别对分离病原的抑菌效果,对7日龄病死肉鸡进行解剖，通过采集死亡肉仔鸡肝脏、脾脏、心脏进行病原菌分离培养、形态观察、16S rDNA分子鉴定，最终鉴定该病死肉仔鸡为大肠杆菌与沙门氏菌混合感染。抑菌结果显示，复方中草药对分离得到的大肠杆菌、鸡白痢沙门氏菌、鸡副伤寒沙门氏菌均有良好的抑菌效果。10种抗生素药敏试验结果显示，此三株菌同时对氨苄西林、环丙沙星、四环素、恩诺沙星四种药物耐药；鸡大肠杆菌对庆大霉素、阿米卡星敏感，鸡白痢沙门氏菌对链霉素、氟苯尼考、头孢噻肟敏感，鸡副伤寒沙门氏菌对头孢曲松、链霉素、头孢噻肟敏感。上述研究结果为肉仔鸡大肠杆菌与沙门氏菌混合感染的分离鉴定以及科学指导用药提供参考。     

用免疫金技术检测猪繁殖与呼吸综合征病毒

用胶体金标记提纯后的兔抗猪繁殖与呼吸综合征病毒 (PRRSV)IgG ,建立了一种以微孔滤膜为固相载体 ,以红色胶体金为标记物的检测PRRSV的斑点免疫金渗滤法 (DIGFA)。特异性阻断试验与交叉试验证明DIGFA检测PRRSV有较高的特异性。检测 1 5份临床样本 ,其中 3份DIG FA及细胞培养均为阳性 ,电镜观察亦可见符合PRRSV特征的病毒粒子     

替米考星与泰乐菌素对畜禽常见病原菌抑菌作用比较

采用试管2倍稀释法对国产替米考星与泰乐菌素进行了畜禽常见的20种病原菌体外抑菌活性比较试验。结果表明,替米考星和泰乐菌素具有类似的抗菌谱,二者对革兰氏阳性菌均有良好的抑制作用。对革兰氏阴性菌二者除对禽多杀性巴氏杆菌、红斑丹毒丝菌、胸膜肺炎放线杆菌和鸡毒支原体均有较强的抑菌效果外,对其他菌则表现为抑菌效果较弱或抗药性。替米考星对部分病原菌如鸡白痢沙门菌、鼠伤寒沙门菌、肺炎克雷伯菌抑菌效果明显强于酒石酸泰乐菌素。总体来看,替米考星对畜禽常见病原菌的抑菌活性比泰乐菌素要强一些。     

Pulsed-field gel electrophoresis genotyping of Salmonella gallinarum and comparison with random amplified polymorphic DNA

Salmonella gallinarum is gram-negative bacteria that cause fowl typhoid (FT) in chickens. Since the first outbreak of FT reported in 1992 in Korea, it has widely spread throughout the country. Today, FT is one of the most devastating diseases of poultry. The aim of the present study was to ascertain a genetic relationship among S. gallinarum isolates collected from different regions of Korea over a 10-year period. We examined a total of 38 isolates of S. gallinarum obtained in 29 regions of Korea from 1992 to 2001 including the 9R vaccine strain and the standard strain of S. gallinarum (ATCC 9184). The PFGE profiles produced 12 different patterns with the XbaI-digestion and 11 different patterns with the SpeI-digestion. The RAPD using URP-6 primers showed eight different genotypes with the same Salmonella isolates. The PFGE patterns of the 9R vaccine strain and ATCC 9184 of S. gallinarum were different from the identical type A, the most common genotype among field isolates in our study. In conclusion, a low genetic heterogeneity was observed among Korean S. gallinarum isolates. In addition, PFGE appeared to be a more accurate and reproducible method for genotyping of S. gallinarum isolates than RAPD.     

Effect of ochratoxin A on broiler chicks challenged with Salmonella gallinarum

1. A study was conducted to evaluate the effects of ochratoxin A (OA) on broiler chicks challenged with Salmonella gallinarum. 2. One hundred and seventy-six 1-d-old broiler chicks were divided into two groups of 88 chicks each, with one group fed on a control mash diet and the other given a mash diet containing 2 ppm OA. On d 14, each group was further subdivided into two groups with one group infected with S. gallinarum and the other uninfected. 3. Following S. gallinarum inoculation on d 14, 4 birds from each group were killed at 1, 2, 3, 5, 7, 10, 14 and 21 d post inoculation. 4. S. gallinarum infection caused dullness, depression, weakness, increased thirst, droopy wings, ruffled feathers and greenish-yellow diarrhoea. S. gallinarum infection in the absence of OA caused 11.5% mortality which increased to 28.8% in the presence of OA. 5. Decreased body weight and reduced feed intake were observed in chicks fed on the diet containing OA. S. gallinarum infection also reduced the body weights of chicks, with the effects being more marked in chicks receiving OA. The OA diet led to increased serum levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, uric acid and creatinine, and decreased levels of total proteins, albumin, globulins, calcium and phosphorus. S. gallinarum infection did not cause significant alteration in any of the serum biochemical parameters. 6. Mortality and the severity of S. gallinarum infection in broiler chicks were increased by the presence of OA in the diet.     

Cell-mediated and humoral immune responses to a virulent plasmid-cured mutant strain of Salmonella enterica serotype gallinarum in broiler chickens

One-week-old Salmonella-free broiler chicks were subcutaneously immunized and subsequently boosted two weeks later with 2x10(7)cfu 0.5ml(-1) of an 85-kb virulent plasmid-cured spectinomycin-resistant mutant strain (SG9VP(-)Spc(r)) of Salmonella gallinarum 9 (S. enterica serotype gallinarum 9) along with a control group of mock-immunized chickens. The chicks were subcutaneously challenged at 5 weeks of age with 5x10(10)cfu 0.5ml(-1) (5xLD(50)) of wild-type S. gallinarum var. Duisburg (S. enterica serotype gallinarum var. Duisburg). The cellular and humoral immune responses were measured at weekly intervals post-immunization (PI) and post-challenge (PC) using lymphocyte stimulation test (LST), delayed type hypersensitivity (DTH) test, serum tube agglutination test (STAT) and enzyme-linked immunosorbent assay (ELISA). High stimulation indices suggestive of a potent lymphocyte transformation response and high persistent serum IgG titres were recorded in immunized chickens at the termination of the experiment. These findings indicate that the live attenuated mutant vaccine induced a strong cellular and humoral immunity, which may play a role in the protection of fowl typhoid in broiler chickens.     

Natural and experimental infection of rodents (Rattus norvegicus) with Salmonella gallinarum]

Thirty five (35) rats (Rattus norvegicus) were trapped in the area of four egg producing poultry farms and were examined for Salmonella spp., micro-biologically. The samples were taken from liver, spleen and intestinal content. Cultures were made directly in MacConkey Agar, in Selenite broth and Rappaport Vassiliadis at 37 degrees C and 43 degrees C. Five strains of S. gallinarum and one strain of Salmonella subgroup II were isolated from the intestinal content of six rats. An experimental study was also carried out. Ten rats Rattus norvegicus were trapped near poultry farms of N. Greece. No Salmonella could be detected in their feces when examined three times by the Selenite 37 degrees C and Rappaport-Vassiliadis 37 degrees C methods. The rats were orally infected with an 18 hours culture of S. gallinarum (1 x 10(9)/ml microorganisms). One hundred and sixty samples of feces were periodically collected and examined for the isolation of the microorganism by the methods mentioned above. Although not any clinical sign of a disease was noticed, S. gallinarum was isolated from their feces up to 121 days post infection.     

壳聚糖对5种细菌体外的抑制试验

为开发利用壳聚糖,选择两种不同性质壳聚糖进行体外抑菌试验。将白色壳聚糖(Ⅰ组)和褐色壳聚糖(Ⅱ组)的乙酸溶液、褐色壳聚糖水溶液(Ⅲ组)用无菌蒸馏水分别稀释至10,5,2.5,1.25 g/L和0.625 g/L,用相应稀释度乙酸溶液(Ⅳ组)作对照组。对大肠埃希菌、金黄色葡萄球菌、蜡样芽孢杆菌、绿脓杆菌、鸡伤寒沙门菌进行体外抑菌试验。结果表明,在体外,壳聚糖对上述5种细菌的生长均有不同程度的抑制作用,不同性质的壳聚糖和同一性质不同浓度的壳聚糖抑菌效果不同。