Use of Fourier-transform infrared spectroscopy to quantify immunoglobulin G concentration and an analysis of the effect of signalment on levels in canine serum     

《Veterinary immunology and immunopathology》2015,163(1-2):8-15

Deficiency in immunoglobulin G (IgG) is associated with an increased susceptibility to infections in humans and animals, and changes in IgG levels occur in many disease states. In companion animals, failure of transfer of passive immunity is uncommonly diagnosed but mortality rates in puppies are high and more than 30% of these deaths are secondary to septicemia. Currently, radial immunodiffusion (RID) and enzyme-linked immunosorbent assays are the most commonly used methods for quantitative measurement of IgG in dogs. In this study, a Fourier-transform infrared spectroscopy (FTIR) assay for canine serum IgG was developed and compared to the RID assay as the reference standard. Basic signalment data and health status of the dogs were also analyzed to determine if they correlated with serum IgG concentrations based on RID results.Serum samples were collected from 207 dogs during routine hematological evaluation, and IgG concentrations determined by RID. The FTIR assay was developed using partial least squares regression analysis and its performance evaluated using RID assay as the reference test. The concordance correlation coefficient was 0.91 for the calibration model data set and 0.85 for the prediction set. A Bland–Altman plot showed a mean difference of −89 mg/dL and no systematic bias. The modified mean coefficient of variation (CV) for RID was 6.67%, and for FTIR was 18.76%.The mean serum IgG concentration using RID was 1943 ± 880 mg/dL based on the 193 dogs with complete signalment and health data. When age class, gender, breed size and disease status were analyzed by multivariable ANOVA, dogs <2 years of age (p = 0.0004) and those classified as diseased (p = 0.03) were found to have significantly lower IgG concentrations than older and healthy dogs, respectively.  相似文献   

Comparison of immune responses to intranasal and intrapulmonary vaccinations with the attenuated Mycoplasma hyopneumoniae 168 strain in pigs     

Pengcheng LI  Yunfeng LI  Guoqing SHAO  Qinghua YU  Qian YANG 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(5):519-525

The aim of this study was to evaluate the immune responses to intranasal and intrapulmonary vaccinations with the attenuated Mycoplasma hyopneumoniae (Mhp) 168 strain in the local respiratory tract in pigs. Twenty-four pigs were randomly divided into 4 groups: an intranasal immunization group, an intrapulmonary immunization group, an intramuscular immunization group and a control group. The levels of local respiratory tract cellular and humoral immune responses were investigated. The levels of interleukin (IL)-6 in the early stage of immunization (P<0.01), local specific secretory IgA (sIgA) in nasal swab samples (P<0.01); and IgA- and IgG-secreting cells in the nasal mucosa and trachea were higher after intranasal vaccination (P<0.01) than in the control group. Interestingly, intrapulmonary immunization induced much stronger immune responses than intranasal immunization. Intrapulmonary immunization also significantly increased the secretion of IL-6 and local specific sIgA and the numbers of IgA- and IgG-secreting cells. The levels of IL-10 and interferon-γ in the nasal swab samples and the numbers of CD4⁺ and CD8⁺ T lymphocytes in the lung and hilar lymph nodes were significantly increased by intrapulmonary immunization compared with those in the control group (P<0.01). These data suggest that intrapulmonary immunization with attenuated Mhp is effective in evoking local cellular and humoral immune responses in the respiratory tract. Intrapulmonary immunization with Mhp may be a promising route for defense against Mhp in pigs.  相似文献   

PRRSV野毒感染猪群免疫接种弱毒活疫苗的效果分析     

许心婷  施开创  李常挺  龙飞翔  莫胜兰  陈汉忠 《中国畜牧兽医》2016,43(9):2447-2454

为评估猪繁殖与呼吸综合征（porcine reproductive and respiratory syndrome,PRRS）弱毒活疫苗的免疫效果,本研究在两个小型猪场各选取两窝仔猪,在免疫接种猪繁殖与呼吸综合征病毒（porcine reproductive and respiratory syndrome virus,PRRSV）弱毒活疫苗后不同时间采集血清,应用RT-PCR检测PRRSV核酸,应用N-ELISA和G-ELISA两种试剂盒检测PRRSV抗体。结果发现,免疫前0 d可以从仔猪血清中检测到PRRSV野毒株核酸,可持续到免疫后30 d;相对应的母猪在免疫前0 d可从血清中检测到PRRSV野毒株核酸。试验仔猪在免疫前0 d未能检测到PRRSV抗体,但在免疫后30～150 d均可检测到PRRSV抗体,其中N-ELISA试剂盒的阳性检出率在免疫后30、60 d高于G-ELISA试剂盒的阳性检出率,而在免疫后120、150 d则低于G-ELISA试剂盒的阳性检出率。使用两种ELISA试剂盒共同检测216份血清样品,检测结果的总体符合率为95.83%;配对χ²检验,P>0.05,两者差异不显著;Kappa值为0.87,属于极强一致性;两者相关系数r为0.605,具有显著线性相关。表明免疫接种弱毒活疫苗可以有效清除野毒感染猪群中的野毒株,N-ELISA和G-ELISA两种ELISA试剂盒均可用于评估弱毒活疫苗的免疫效果。  相似文献   

油菜菌核病菌致病性分化及弱毒现象     

邓小军  李桂芳  魏颖  庞茜丹  朱宏建  高必达 《湖南农业大学学报(自然科学版)》2016,42(3):317-321

对湖南省常德、益阳两地的196个油菜菌核病菌株进行分离纯化,采用离体叶片法测定它们的致病力,发现两地区油菜菌核病菌的致病力明显分化。运用纤维素吸附法提取19个弱致病力菌的ds RNA,其中3个菌株含有ds RNA,菌株CY019菌丝稀疏,菌核量少。进一步利用尖端脱毒获得不含ds RNA的菌株CY019VF,通过活体茎秆接种,发现其致病力得到恢复,表明CY019致病力下降可能与其所携带的ds RNA病毒有关。  相似文献   

Immune Responses to the Attenuated Mycoplasma hyopneumoniae 168 Strain Vaccine by Intrapulmonic Immunization in Piglets   总被引：6，自引：0，他引：6  

FENG Zhi-xin  SHAO Guo-qing  LIU Mao-jun  WU Xu-su  ZHOU Yong-qi  GAN Yuan 《中国农业科学(英文版)》2010,9(3):423-431

To investigate the immune responses to the attenuated Mycoplasma hyopneumoniae 168 strain vaccine, 8-15 d old piglets were immunized with M. hyopneurnoniae 168 strain vaccine by intrapulmonic route. And the specific IgG antibody in serum, lymphoproliferation, IFNT, and specific secretory IgA （SIgA） antibody in bronchoalveolar lavage fluid were detected on 30 and 60 d post-immunization （DPI）, respectively. On 60 DPI, all the pigs except for those in health control group were challenged with a field M. hyopneumoniae strain JS. Necropsy was performed on 30 d post-challenge （DPC）. The results showed that IFN7 and specific SIgA were stimulated on surface of respiratory tract after immunization. And peripheral blood mononuclear cells could also be proliferated about 1.81 and 2.12 fold on 30 and 60 DPI when stimulated by M. hyopneumoniae protein in vitro. However, no serum IgG antibody against M. hyopneumoniae was detected during the whole immune phage. After challenge, vaccinated pigs were observed with only very slight histological lesion in individual lobes. None of vaccinated pigs showed any clinical signs. While the unvaccinated pigs from challenge control group showed varying degrees of clinical sign and severe macroscopical lesion of mycoplasmal pneumonia of swine （MPS）. The result suggested that the attenuated M. hyopneumoniae 168 strain vaccine inoculated by intrapulmonic route could activate the systemic cellular immunity, the local mucosal immunity and IFNγ secretion in respiratory tract to against M. hyopneumoniae infection in piglets.  相似文献   

检测鸡球虫四价弱毒疫苗抗体应答的ELISA方法的建立及初步应用   总被引：1，自引：0，他引：1  

李仁良  袁子国  侯杰  谭志坚  林瑞庆  朱兴全  翁亚彪 《中国畜牧兽医》2011,38(8):52-55

本研究旨在建立检测鸡球虫四价弱毒疫苗(柔嫩艾美耳球虫、毒害艾美耳球虫、堆型艾美耳球虫、巨型艾美耳球虫)抗体应答的酶联免疫吸附试验(ELISA),并初步阐明鸡免疫四价弱毒疫苗及攻虫后的抗体应答。将300羽岭南黄鸡分为A(免疫攻虫组)、B(不免疫不攻虫组)、C(不免疫攻虫组)共3个组。A组雏鸡在4日龄时进行首免,11日龄时鸡开始二免,二免完成后(17日龄时)对A组及C组的鸡进行攻虫(28万个4种球虫的混合卵囊/羽),7 d后检查A、C两组鸡的存活率。在鸡二免和三免完成后,分别对A、B组进行采血,通过优化最佳抗原包被浓度、酶结合物工作浓度及最佳血清稀释度,建立了ELISA方法来检测鸡体对球虫四价弱毒疫苗的抗体应答情况。A组鸡血清抗体的平均D值为0.870和0.904,明显高于B组鸡的平均D值0.261和0.270,证明了鸡体免疫疫苗后可以刺激产生相应抗体。免疫鸡攻虫后的存活率和抗体应答的检测结果,都充分说明了此鸡球虫弱毒四价疫苗具有良好的免疫原性,所建立的ELISA方法为今后评价鸡球虫弱毒四价疫苗的免疫效力提供了有效手段。  相似文献   

鸡传染性法氏囊病分子流行病学研究     

白文霞  鲍恩东  侯继波  何孔旺 《猪业科学》2004,21(8):51-53

15个江苏省IBDV分离株,对其VP2高变区进行RT-PCR扩增、测序,得到约560bp长的片段。分析比较15个IBDV分离株与参考毒株的VP2高变区(AccI-SpeI)推断的氨基酸序列,构建进化树。氨基酸序列分析以及进化树分析表明,15个IBDV分离株是超强毒株。而且目前vvIBDVs在亚洲各个国家流行,且与欧洲国家的vvIBDVs的进化关系很近。  相似文献   

不同剂量CpG序列对仔猪的免疫增强效果   总被引：7，自引：0，他引：7  

田兴山张玲华  邝哲师陈薇 李健雄 《华南农业大学学报》2004,25(1):96-98

以伪狂犬病毒为模式病毒,按照仔猪体质量分别将100、10、1μg/kg CpG序列同猪伪狂犬疫苗联合免疫接种仔猪,检测仔猪的体液及细胞免疫反应．试验结果表明：不同浓度的CpG序列均能显著提高仔猪的特异性抗体滴度、白细胞介素(IL)-2诱生活性以及淋巴细胞增殖反应,证明CpG序列能显著增强仔猪对常规疫苗的免疫应答能力。  相似文献   

小鼠对pGM-CSF/SS生长抑素基因疫苗的免疫应答     

梁爱心  姚艳丰  金定恩  武睿  韩丽  付平  刘耘  杨利国 《华中农业大学学报》2006,25(5):540-543

通过研究细胞因子GM-CSF与生长抑素pcS/2SS融合表达质粒pGM-CSF/SS诱导小鼠免疫应答的影响,探求生长抑素基因免疫新策略。将60只20日龄雄性昆明小白鼠均分为6组,分别用PBS(C0)、pGM-CSF质粒(C1)、pGM-CSF/SS质粒(T1、T2、T3)、pcS/2SS(T4)质粒口服免疫,T1~T4组的免疫剂量分别为2×106cfu/mL、2×108cfu/mL、2×1010cfu/mL、2×108cfu/mL,以上质粒均以减毒沙门氏菌为传导载体。结果显示:pGM-CSF/SS组的抗生长抑素抗体AP/AN值显著高于pcS/2SS组,阳性鼠体增重亦高于pcS/2SS组,抗体阳性率分别是71.4%和40.0%。另外,高剂量组的AP/AN值极显著高于低剂量组,显著高于中剂量组;中剂量组的AP/AN值显著高于低剂量组,高剂量组的抗体阳性率也明显高于中剂量组和低剂量组,分别为100%、71.4%和20.0%。可见,pGM-CSF/SS疫苗的免疫效果优于pcS/2SS疫苗,且pGM-CSF/SS疫苗呈剂量依赖关系。这些结果显示出细胞因子GM-CSF可以增强生长抑素DNA疫苗的免疫效果。  相似文献   

减毒沙门氏菌介导H9亚型禽流感DNA疫苗与灭活疫苗联合应用研究   总被引：1，自引：0，他引：1  

潘志明  程宁宁  刘蓓蓓  孙林  耿士忠  陈祥  黄金林  焦新安 《扬州大学学报(农业与生命科学版)》2007,28(4):1-5

构建获得含有H9亚型禽流感病毒血凝素(HA)基因的重组真核表达质粒pCAGGS-HA。重组质粒电转化减毒鼠伤寒沙门氏菌SL7207,获得重组沙门氏菌SL7207(pCAGGS-HA)。重组菌SL7207(pCAGGS-HA)以5×109cfu的剂量2次口服免疫1日龄商品代伊莎褐蛋鸡后,再以油乳剂灭活疫苗加强免疫1次,并设立重组菌单独免疫组、灭活苗免疫组、空载体免疫组及空白对照组。联合免疫组和重组菌单独免疫组的小肠黏膜抗体效价与其他组之间存在显著差异(P<0.05),且两组的攻毒免疫保护水平与空载体组之间差异显著(P<0.05),其中联合免疫组的免疫保护率最高,达100%,而灭活苗免疫组保护率为80%,表明减毒沙门氏菌介导的H9亚型禽流感DNA疫苗与灭活疫苗联合应用具有良好的免疫协同作用。  相似文献