鹦鹉热衣原体real-time quantitative PCR检测方法的研究   总被引：1，自引：0，他引：1  

吴东海  李应国  杨迎伍  邓伟  王昱  李正国 《安徽农业科学》2008,36(23)

[目的]为鹦鹉热衣原体的快速、准确检测奠定基础。[方法]根据鹦鹉热衣原体主要外膜蛋白(MOMP)基因序列设计一对特异性引物,以SYBR GreenⅠ为荧光染料,建立了鹦鹉热衣原体Real-time quantitative PCR检测方法。根据检测结果,计算样品的批内和批间变异系数(CV)。[结果]以提取的鹦鹉热衣原体DNA为模板,用引物Cps-1和Cps-2进行PCR扩增,得到长100 bp的片段。扩增产物回收纯化后,连接到pGEM-T Easy载体上并转化到大肠杆菌DH5α,菌落PCR检测筛选阳性克隆,培养后提取质粒DNA。当模板浓度范围为1.78×102~1.78×108拷贝/μl时,标准曲线相关系数达0.998;批内和批间变异系数(CV%)分别为1.30%~4.59%和5.72%~9.87%。[结论]为鹦鹉热衣原体的感染、流行调查等提供了重要的技术参考。  相似文献   

海洋贝类细胞内寄生原核生物研究进展   总被引：1，自引：0，他引：1  

刘英杰  潘金培 《中国水产科学》2004,11(2):170-176

在海洋贝类中寄生的细胞内寄生原核生物主要包括立克次体(Rickettsia)和农原体(Chlamydia),通常寄生于宿主的鳃和消化腺上皮组织细胞的细胞质内,一般形成嗜碱性包涵体,可对宿主细胞造成一定的损伤,但在一般情况下不会引起宿主的系统病理变化,也不会导致死亡。本文对该方面国内外研究的历史、进展和现状进行了系统的总结,对海洋贝类原核生物感染的宿主范围、危害、检测方法及存在的问题进行了阐述,对该领域今后的发展方向进行了展望。  相似文献   

Chlamydia pneumoniae infection induces vascular smooth muscle cell migration through stimulating Rac1 activity via PI3K activation     

ZHANG Jun-xia  ZHANG Teng-teng  ZHANG Li-jun  WANG Bei-bei  WEI Jun-yan  WANG Hai-wei  SHEN Bing-ling  ZHANG Li-jun 《园艺学报》2013,29(2):236-241

AIM: To determine the role of Rac1 activation in the migration of vascular smooth muscle cells (VSMCs) induced by Chlamydia pneumoniae (C.pn) infection and to investigate the effect of phosphatidylinositol 3-kinase (PI3K) on Rac1 activation during the process. METHODS: The recombinant plasmid encoding glutathione S-transferase (GST)-p21-binding domain of p21-activated kinase 1(PBD) fusion protein was transformed into the competent bacteria to induce the expression of the fusion protein. The fusion protein was purified. GST-pull down assay was performed to evaluate the activity of Rac1 in C.pn-infected VSMCs pretreated with or without PI3K inhibitor LY294002 (25 μmol/L). Wound-healing assay and Transwell assay were performed to observe the changes of C.pn infection-induced VSMCs migration with or without pre-incubation of Rac1 inhibitor NSC23766 (50 μmol/L). RESULTS: Enough and biologically active GST-PBD fusion protein was obtained after purification. The activity of Rac1 in VSMCs infected with C.pn for 24 h increased significantly and was higher than that in control group. Rac1 activation induced by C.pn infection was inhibited by the pretreatment of VSMCs with LY294002. The migration ability of C.pn-infected VSMCs pre-incubated with NSC23766 was significantly reduced and was lower than that in C.pn infection group. CONCLUSION: C.pn infection induces the migration of VSMCs possibly through stimulating Rac1 activity via PI3K activation.  相似文献   

布鲁氏菌和鹦鹉热衣原体双重PCR检测方法的建立与应用     

彭武丽  罗展荣  史茜  员丽娟  于学辉  季新成 《中国畜牧兽医》2014,41(4):7-11

为建立同时检测布鲁氏菌和鹦鹉热衣原体的双重PCR方法,本研究据GenBank上已发表的具有属间特异性的布鲁氏菌bp26基因和鹦鹉热衣原体23S rRNA基因,利用 Primer Premier 5.0软件各设计1对特异性引物,扩增的目的片段长度分别为219和356 bp。通过优化反应条件,建立了能同时检测布鲁氏菌和鹦鹉热衣原体的双重PCR方法。该方法具有较好的特异性和可重复性,对2种基因单重PCR检测敏感性均达到3.1×10²拷贝/反应,双重检测的灵敏度为3.1×10³拷贝/反应。利用该双重PCR方法对流产牛抗凝全血、血清、流产胎儿及奶液共172份临床疑似布鲁氏菌感染的样品进行检测,检测到布鲁氏菌阳性样品53份,鹦鹉热衣原体阳性样品2份,以上这2种病原的阳性检出率分别为30.8%和1.2%,且检测到2种病原混合感染的阳性样品2份,阳性检出率为1.2%。临床应用结果表明,该方法可用来对布鲁氏菌和鹦鹉热衣原体进行同步、快速、灵敏的检测。  相似文献   

Comparison of the omp I gene of Chlamydia psittaci between isolates in Victorian koalas and other animal species     

JL MARTIN  GF CROSS 《Australian veterinary journal》1997,75(8):579-582

Objective The objective of this study is to compare the strain of chlamydia causing genital infection in koalas from Victoria with isolates from other animal species.
Design Polymerase chain reaction and restriction enzyme analysis has been used to compare various Chlamydia psittaci isolates from a range of animals and disease syndromes. The isolates used in this study include isolates from three birds, three from aborted sheep, one from polyarthritis, one from bovine abortion, one from feline pneumonitis, three porcine isolates from faeces, polyarthritis and abortion, and three urogenital isolates from Victorian koalas.
Procedure Two polymerase chain reactions were performed, each amplifying a different region of the omp I gene. The first polymerase chain reaction amplified a 144 bp segment of the gene which was then digested with the restriction enzyme Eco R I. The second polymerase chain reaction amplified a larger 1070 bp region of the omp I gene which was digested with two restriction enzymes Alu I and Nde II.
Results and conclusions The results obtained have confirmed that variation in DNA sequence of various animal chlamydia isolates does occur. They have also shown that it is possible to classify isolates, based on their restriction enzyme profiles, into distinct groups.  相似文献   

免疫萤光技术检测禽用活疫苗中衣原体的污染     

吴福林 《中国兽药杂志》1997,31(1):17-19

本文主要描术字用萤光抗体技术对４０批禽病疫苗，以不同的形式进行了检测。第一，取１０批新城疫疫苗，分别用ＮＤ阳性血清中和，接种鸡胚，并任取一批与１０＾－４稀释的衣原体混合后接种鸡胚，作为阳性对照；第二，取２０批禽病疫苗直接涂片检测同时设衣原体对照；第三，取１０批ＮＤ苗，用ＮＤ阳性血清中和，接种鸡胚，同时每批苗分别为１０＾－１４衣原体稀释液混合，接种鸡胚作对照，结果４０批疫苗均未检出阳性，衣原体对照均  相似文献   

禽源鹦鹉热衣原体MOMP基因重组腺病毒载体的构建与鉴定   总被引：1，自引：0，他引：1  

《中国兽医科技》2006,36(1):13-17

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Genetic and antigenic analysis of Chlamydia pecorum strains isolated from calves with diarrhea     

Akifumi OHTANI  Masahito KUBO  Hiroshi SHIMODA  Kenji OHYA  Tadashi IRIBE  Daiki OHISHI  Daiji ENDOH  Tsutomu OMATSU  Tetsuya MIZUTANI  Hideto FUKUSHI  Ken MAEDA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(7):777-782

Chlamydia pecorum (designated 22–58) was isolated in 2010 in HmLu-1 cells from the jejunum of a calf which died of necrotizing enterocolitis in Yamaguchi Prefecture, Japan. Immunohistochemical staining identified C. pecorum positive reactions in the jejunal villi. C. pecorum, designated 24–100, was isolated from the feces of a calf with diarrhea in another farm in Yamaguchi Prefecture in 2012. A significant increase in neutralizing antibody titers against C. pecorum was confirmed in paired sera. Nucleotide sequence identities of omp1 genes of the 2 isolates were 100%. The isolates were genetically and antigenically more closely related to C. pecorum Bo/Yokohama strain isolated from cattle with enteritis in Japan than to the other prototype strains, Bo/Maeda isolated from cattle with pneumonia and Ov/IPA isolated from sheep with polyarthritis. These results indicate that C. pecorum strains similar to 22–58 and 24–100 might be endemic in Yamaguchi Prefecture and cause enteric disease in cattle.  相似文献   

山东省部分地区家禽鹦鹉热衣原体感染情况调查     

李婧  王梅  刘新勃  李兆华  李舫 《中国预防兽医学报》2015,37(7)

为了解山东地区家禽鹦鹉热衣原体(Cps)感染现状,本研究采用间接血凝法(IHA)对2013年~2014年采集自山东潍坊、淄博、济南、临沂、烟台等地区的1 020份鸡、鸭血清样品进行Cps抗体的检测,并对检测数据进行了统计分析;结果显示:IHA测得总阳性率为29.51%(301/1 020);鸡阳性率为25.26%(197/780);鸭阳性率为43.33%(104/240);各地区间阳性率存在一定差异。本调查结果表明,家禽Cps感染在山东地区具有较高的感染率。  相似文献   

Seroprevalence Investigation and Risk Factors Analysis of Chlamydia Infection in Dairy Cattle in Gansu and Ningxia Areas     

TAN Qi-dong  LI Zhi-xin  WANG Xiao-liang  YIN Ming-yang  QIN Si-yuan  LIU Guang-xue  ZHU Xing-quan  ZHOU Dong-hui  XU Qian-ming 《中国畜牧兽医》2015,42(5):1283-1287

The objective of the present investigation was to examine seroprevalence of Chlamydia in dairy cattle in Gansu and Ningxia areas, Northwest China, and to analyze the factors affecting Chlamydia infection in dairy cattle.Indirect hemagglutination assay (IHA) was used to detect antibodies against Chlamydia in 1 657 dairy cattle serum samples from Gansu and Ningxia areas.Epidemiological investigation and statistical methods were used to analyze the data.The results showed that the overall seroprevalence was 29.33%, logistic regression analysis was applied to evaluate the risk factors of Chlamydia infection in dairy cattle, and the results indicated that age and numbers of pregnancy of dairy cattle were not the significant risk factors (P>0.05), and were left out of the final model, however, region was considered as the main risk factor associated with Chlamydia infection (P<0.05).The highest titer was 1:1 024.In conclusion, the results of the present survey indicated the widespread of Chlamydia infection in Gansu and Ningxia areas.In order to ensure the economic benefit of dairy farming, we should pay more attention to Chlamydia infection in dairy cattle, and integrated control strategies and efficient management measures should be implemented to prevent and control Chlamydia infection in dairy cattle.  相似文献