Evaluation of PCR,IEF and ELISA techniques for the detection and identification of potato cyst nematodes from field soil samples in England and Wales     

Said K Ibrahim  Stephen T Minnis  Anthony D P Barker  Mike D Russell  Patrick P J Haydock  Kenneth Evans  Ivan G Grove  Simon R Woods  Andrew Wilcox 《Pest management science》2001,57(11):1068-1074

Effective management of potato cyst nematodes (PCNs) requires simple, rapid and accurate identification and quantification of field populations. Soil samples from a survey of 484 fields in potato rotations in England and Wales were used to compare the identification and quantification of PCNs using IEF, PCR, ELISA and bait plant tests. The cyst counts and bait plant test revealed that 64.3% of field samples contained PCNs. Bait plant tests increased the detection rate of PCNs in field samples by 4–6.4%. This means that some infestations are cryptic and would not normally be detected by standard counts. IEF, PCR and ELISA methods distinguished between Globodera rostochiensis and G pallida and were able to register mixed populations; however they were not in full agreement. All methods suggested that G pallida is the dominant species in the field samples tested. The PCR results indicated that 66% of field samples contained pure G pallida, 8% contained pure G rostochiensis and 26% contained mixtures of the two species. Estimates of the relative process times taken per sample in the PCR, IEF and ELISA techniques are given. © 2001 Society of Chemical Industry  相似文献   

7种长蠹科昆虫的线粒体DNA ND4基因序列比较分析   总被引：6，自引：0，他引：6  

李伟丰  黄永成  陈邦禄  陈永久  张亚平 《植物检疫》2001,15(5):257-262

长蠹科昆虫严重危害林木和仓贮物品。该文应用非损伤性DNA测序技术测定了来自不同国家的长蠹科害虫的线粒体DNA ND4基因的部分序列。在获得的204bp的序列中，7种昆虫的序列变异丰富，多数变异发生在密码子的第3位点上。将实验结果与形态学特征比较分析，探讨7个种的形态差异与基因序列的差异；结果表明，7种昆虫不仅在外形特征上存在差异，而且在ND4基因序列上的差异程度也很显著，平均为21．94％。这为分类鉴定提供了充分的证据。  相似文献   

Interaction of benzimidazole-N-sulfonamides with the cytochrome b/c1 complex of Pythium aphanidermatum     

Christian Pillonel 《Pest management science》1995,43(2):107-113

Experiments with intact cells and submitochondrial fractions of Pythium aphanidermatum (Edson) Fitz. indicated an interference of benzimidazole-N-sulfonamides with the NADH- or succinate-driven electron transport system between cytochromes b and c. Comparison with Ustilago maydis (DC) Corda and Botrytis cinerea Pers. ex Fr. revealed that this effect is Oomycetes specific. The molecular interaction between benzimidazole-N-sulfonamides and the mitochondrial cytochrome b/c₁ complex from P. aphanidermatum has been investigated. Binding assays with [¹⁴C]52232 RP (dimefluazole) indicated a time- and dose-dependent labelling of two proteins. The molecular mass of one labelled protein and the competition of the binding with antimycin A suggest that benzimidazole-N-sulfonamides interact with the Q₁-centre of cytochrome b. Furthermore, experiments with doubly labelled [³H][¹⁴C]CGA 323103 revealed a possible irreversible inactivation of the b/c₁ complex leading to covalent linkage of the dimethylsulfonamoyl moiety to the target site.  相似文献   

缓变假说     

韩田夫  陈维多 《东北农业大学学报》1991,(3)

本文首次提出缓变的概念。缓变是因 DNA 高级结构变化形成新的基因序列而引起的变异。作者认为,缓变有一定的分子基础,可以解释适应、分化和获得性状遗传等现象,并指出基因在碱基的编码序列(DNA 一级结构)上可以是不连续的。但在 DNA 高级结构中是连续的。DNA 负载遗传信息的能力不仅限于碱基序列,而且包括空间结构。  相似文献   

安哥拉山羊与建昌黑山羊及其杂种后代的DNA指纹分析   总被引：5，自引：0，他引：5  

王杰  沈富军  欧阳熙  王永  覃志红 《中国畜牧杂志》2001,37(3):13-15

以 (CA/GATA/TCC) 5探针 ,HinfI酶切 ,研究安哥拉山羊和建昌黑山羊及安哥拉山羊与建昌黑山羊级进杂交的F2 、F3的DNA指纹图谱。结果 ,在供试山羊中 ,个体平均检出 1 8.2± 0 .4条谱带 ,安哥拉山羊、F2 、F3的鉴别机率分别为 1 .38× 1 0 - 12 ,1 .1 8× 1 0 - 13,0 .50× 1 0 - 10 ;群体内相似系数安哥拉山羊为0 .4 539,F2 为 0 .4 0 77,F3为 0 .61 1 1 ;亲子鉴定的父权概率W =0 .9889;安哥拉山羊与建昌黑山羊比较 ,2 .3kb和 8.6kb谱带为安哥拉山羊的特异谱带  相似文献   

Detection of papillomavirus-DNA in mesenchymal tumour cells and not in the hyperplastic epithelium of feline sarcoids     

Teifke JP  Kidney BA  Löhr CV  Yager JA 《Veterinary dermatology》2003,14(1):47-56

Abstract We examined 12 formalin-fixed paraffin-embedded feline skin tumours which had the histopathological features of fibropapillomas for the presence of papillomavirus (PV) DNA using touchdown polymerase chain recation (PCR), DNA sequencing and nonradioactive in situ hybridization. Nine of the tumours contained a 102-bp PCR product demonstrated using consensus PV primers that amplify a portion of the L1 gene. The nucleotide sequences are closely related, but not identical to that of ovine PV type 2, rabbit oral PV and reindeer PV. The deduced amino acid sequences had strong homologies with the major capsid protein L1 of deer PV, bovine papillomavirus (BPV) 1 and BPV 2, and European elk PV. Although PV antigens were not detected in any of the tumours by immunohistochemistry, PV DNA was demonstrated in individual mesenchymal cells or cell nests of 4/12 tumours by in situ hybridization. A nonproductive infection of mesenchymal fibroblast-like tumour cells with a papillomavirus would explain the lack of PV antigen expression and the absence of PV DNA in the hyperplastic epithelium. Because these tumours and their pathogenesis are similar to equine sarcoids, we suggest that they should be reclassified as 'feline sarcoids' instead of fibropapillomas.  相似文献   

The loss of genetic diversity in Dabry's sturgeon (Acipenser dabryanus,Dumeril) as revealed by DNA fingerprinting     

Qiu‐Hong Wan  Sheng‐Guo Fang  Ya‐Nan Li 《水产资源保护:海洋与淡水生态系统》2003,13(3):225-231

• 1. Dabry's sturgeon, a large, long‐lived migratory fish is endemic to the Yangtze River. Over‐fishing and habitat destruction have caused large‐scale declines in natural stocks in the last two decades.
• 2. Examining patterns of genetic diversity has become an integral component of many management plans for endangered species. DNA fingerprinting was applied to detect genetic diversity in Dabry's sturgeon collected in 1958–1959, 1980–1981 and 1998–1999.
• 3. Studies on direct genetic parameters (genetic variability, hypervariable loci and heterozygosity) and indirect parameters (band‐sharing coefficient and allelic frequency) showed that the continuous decline in wild populations has caused the loss of genetic diversity in present‐day sturgeon.
• 4. The present‐day populations have the lowest genetic variability; thus, effective management is needed to preserve genetic diversity.
• 5. A conservation strategy is urgently required, comprising artificial rearing facilities coupled with breeding management plans.

Copyright © 2003 John Wiley & Sons, Ltd.  相似文献   

New gene-based approaches for an AIDS vaccine     

Puaux AL  Michel ML 《Comparative immunology, microbiology and infectious diseases》2003,26(5-6):357-372

Vaccine approaches against AIDS have focused on inducing cellular immune responses, since many studies revealed the role of T cell responses in the control of human immunodeficiency virus or simian immunodeficiency virus (SIV) infections. The experimental infection of rhesus macaques with SIV or chimeric SHIV is routinely used as a model for AIDS. In such models, DNA immunization is a tool to elicit specific T cell responses and to study their protective efficacy. DNA immunogenicity in primates depends on parameters such as level of antigen expression, choice of the antigen among SIV proteins, use of fusion proteins, route of immunization, and addition of adjuvants. Recent results suggest that priming with DNA and boosting with attenuated recombinant viral vectors, each expressing corresponding SIV antigens, leads to improved specific immunity and, in some cases, affords protection against pathogenic challenge. After preclinical evaluations, DNA has entered clinical trials for a therapeutic or prophylactic gene-based AIDS vaccine.  相似文献   

Flow cytometry: clinical applications in equine medicine     

Davis EG  Wilkerson MJ  Rush BR 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2002,16(4):404-410

The use of flow cytometry in veterinary diagnostics is becoming a valuable clinical tool with a broad range of applications. Physical characteristics of cells can be determined by the flow cytometer laser and electronics through the measurement of changes in light scatter properties. Other components and functions of cells can be defined through the application of fluorochrome dyes that have an affinity for cellular components. Traditionally, common clinical applications are immunophenotyping of cells of the hematopoietic system with fluorescent-labeled antibodies raised against specific cell surface proteins. Other approaches have been used to elucidate changes in cell function and DNA content. This review is intended to provide the reader with the fundamental uses of flow cytometry. Examples of clinical applications in equine patients include immune-mediated hemolytic anemia, immune-mediated thrombocytopenia (IMT), chronic inflammatory disease, and neoplasia.  相似文献   

Evaluation of the Abbott LCx Mycobacterium Tuberculosis Assay for Direct Detection of Mycobacterium Bovis in Bovine Tissue Samples     

Valente C  Cuteri V  Ausili E  Piersimoni C 《Veterinary research communications》2002,26(1):21-27

The commercial LCx amplification assay, usually employed to detect the Myocobacterium tuberculosis complex in respiratory specimens, was evaluated by comparing the results it gave with those obtained using Löwenstein-Jensen solid medium and pathological findings on 55 lymph nodes from cattle with positive and 10 lymph nodes from cattle with negative skin tests for tuberculosis. Fifty-three cultures (51 and 2, respectively) were positive for M. bovis, while the results for the LCx assay and the histological method were positive in 48 (45, 3) and 24 (20, 4) samples, respectively. None of the samples from cattle from certified tuberculosis-free herds were positive by any of the procedures. The results obtained with the LCx assay, compared with the culture procedure, regarded as the gold standard among the diagnostic techniques, gave a specificity of 91.6% and sensitivity of 90.5%. Although the sensitivity of LCx was suboptimal, DNA of M. bovis was detected in 81.8% of the skin test-positive animals. Amplification techniques could provide a rapid and reasonably reliable tool for detecting bovine tuberculosis.  相似文献