Liman M.,AniCon Labor GmbH |
Block J.,AniCon Labor GmbH |
Hellmers N.,AniCon Labor GmbH |
Wolking D.,AniCon Labor GmbH |
Behr K.-P.,AniCon Labor GmbH
European Poultry Science | Year: 2014
The introduction of biomolecular methods into routine veterinary diagnostic work flow for the poultry industry dramatically changed and improved the overall diagnostic possibilities, especially with regards to fast detection of viruses. The biomolecular methods based on amplification techniques such as Polymerase Chain Reaction and its derivatives and on subsequent or real-time detection, were extensively implemented in poultry veterinary diagnostics in the last three decades. Today all essential components, such as PCR thermal cycler, software, detection kits and on-site training, needed to establish an accredited lab routine for modern molecular methods, e.g. in an integrated poultry producing company, are available. Other biomolecular methods, such as those based on isothermal amplification, feature significant advantages over Polymerase Chain Reaction and its derivates and may be of importance in the future, but were not recognized by routine veterinary diagnosticians adequately, yet.An overview of published, biomolecular methods either currently applied or of future interest for pathogens relevant for poultry species is given and the present application for major pathogens is described. The requirements for implementation of a modern molecular lab for routine pathogen detection are presented and benefits as well as drawbacks of biomolecular methods are discussed. © Verlag Eugen Ulmer, Stuttgart.
Molecular typing of Riemerella anatipestifer serotype 14, an emerging pathogen for ducks [Zur molekularen Typisierung von feldstämmen von Riemerella anatipestifer serotyp 14, einem Krankheitserreger mit zunehmender Bedeutung für Enten]
Philipp H.-C.,Lohmann Animal Health GmbH |
Taras D.,Lohmann Animal Health GmbH |
Liman M.,Anicon Labor GmbH |
Grosse-Herrenthey A.,Lohmann Animal Health GmbH |
And 2 more authors.
Archiv fur Geflugelkunde | Year: 2013
Riemerella anatipestifer (R. anatipestifer) causes economically important infections in waterfowl and turkeys. Autogenous vaccines are widely used in duck farming due to the high antigenic variablity of R. anatipestifer and the lack of commercial vaccines. Since a few years, we have isolated increasing numbers of R. anatipestifer serotype 14 from vaccinated and non-vaccinated ducks. To further investigate the epidemiology and to improve the efficacy of autogenous vaccines, we used 49 R. anatipestifer serotype 14 strains and two controls belonging to other serotypes for further typing by pulse-field gel electrophoresis (PFGE) and matrix- assisted laser desorption-ionisation time-of-flight mass spectrometry (MALDI-TOF). The strains were isolated between 2008 and 2012 from four different geographic regions, all from ducks with the exception of one isolate originating from a goose. Seven distinct PFGE-patterns forming three clusters were obtained. Group A1 contained all 4 isolates from one of the regions. Group A contained 32 isolates from northwest Germany detected from 2010 onwards suggesting a recent emergence of this clone. All isolates in the study were correctly identified as R. anatipestifer by MALDI-TOF MS. The serotype 14 strains grouped into 8 closely related clusters by using the standard MALDITOF MS software. The clusters were unrelated to the data obtained by PFGE indicating that the currently available standard procedures of MALDI-TOF MS are suitable for the species identification of R. anatipestifer, but not for further subtyping within a serogroup. However, both methods confirm that there is considerable variability between R. anatipestifer isolations even within the same serotype. The genotyping by PFGE provides epidemiological information and identifies potentially emerging clones of R. anatipestifer. Such results are of particular value for the production of efficacious autogenous vaccines. © Verlag Eugen Ulmer, Stuttgart.
Gasch C.,University of Hamburg |
Gasch C.,Queensland University of Technology |
Oldopp T.,University of Hamburg |
Oldopp T.,Anicon Labor GmbH |
And 9 more authors.
Molecular Oncology | Year: 2016
Modern technologies enable detection and characterization of circulating tumor cells (CTC) in peripheral blood samples. Thus, CTC have attracted interest as markers for therapeutic response in breast cancer. First studies have incorporated CTC analyses to guide therapeutic interventions and stratification of breast cancer patients. Aim of this study was to analyze characteristic features of CTC as biomarker for predicting resistance to HER2-targeted therapies. Therefore, CTC from metastatic breast cancer patients with HER2-negative primary tumors screened for the prospective randomized phase III trial DETECT III were explored for their HER2 status and the presence of PIK3CA mutations. Detection and characterization of HER2 expression of CTC were conducted with the CellSearch® system. Fifteen of 179 CTC-positive patients (8.4%) contained ≥1 CTC with strong HER2 expression. Genomic DNA from individual CTC isolated by micromanipulation was propagated by whole genome amplification and analyzed for PIK3CA mutations in exons 9 and 20 by Sanger sequencing. One or more CTC/7.5 mL were detected in 179/290 patients (61.7%). In 109 patients (34.8%), ≥5 CTC/7.5 mL were found. We detected at least one CTC with the mutation p.E542K, p.E545K, p.H1047R, p.H1047L or p.M1043V in 12/33 patients (36.4%). Thirty six of 114 CTC (31.6%) harbored one of these mutations. CTC in individual patients exhibited heterogeneity concerning PIK3CA mutations and HER2 expression. In conclusion, clinically relevant genomic aberrations such as mutations in the hotspot regions of exon 9 and 20 of the PIK3CA gene can be detected in single CTC and might provide insights into mechanisms of resistance to HER2-targeted therapies. © 2016 Federation of European Biochemical Societies
Negash T.,University of Veterinary Medicine Hannover |
Liman M.,University of Veterinary Medicine Hannover |
Liman M.,AniCon Labor GmbH |
Rautenschlein S.,University of Veterinary Medicine Hannover
Vaccine | Year: 2013
Infectious bursal disease virus (IBDV) is an immunosuppressive virus of chickens. The virus protein (VP) 2 induces neutralizing antibodies, which protect chickens against the disease. The aim of this study was to develop a cationic poly(d,. l-lactide-co-glycolide) (PLGA) microparticle (MP) based IBDV-VP2 DNA vaccine (MP-IBDV-DNA) for chickens to be delivered orally and by eye drop route. The tested IBDV-VP2 DNA vaccines were immunogenic for specific-pathogen-free chickens and induced an antibody response after intramuscular application. Co-inoculation with a plasmid encoding chicken IL-2 (chIL-2) or CpG-ODN did not significantly improve protection against IBDV challenge. However, the application of a MP-IBDV-DNA vaccine alone or in combination with a delayed oral and eye drop application of cationic MP loaded with CpG-ODN or chIL-2 improved protection against challenge. The MP-IBDV-DNA-vaccinated chickens showed less pathological and histopathological bursal lesions, a reduced IBDV antigen load as well as T-cell influx into the bursa of Fabricius (BF) compared to the other groups (p< 0.05). The addition of chIL-2 loaded MP improved challenge virus clearance from the BF as demonstrated by lower neutralizing antibody titers and reduced IL-4 and IFN-α mRNA expression in the bursa at 7 days postchallenge compared to the other challenged groups. Overall, the efficacy of the IBDV-DNA vaccine was improved by adsorption of the DNA vaccine onto cationic PLGA-MP, which also allowed mucosal application of the DNA vaccine. © 2013 Elsevier Ltd.