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Acta oncologica. American journal of epidemiology. Correction factors for self-selection when evaluating screening programmes. Journal of medical screening. Contributing factors and outcomes of treatment refusal in pediatric oncology in Germany. Home pesticide exposures and risk of childhood leukemia: Findings from the childhood leukemia international consortium. International journal of cancer Journal international du cancer. Family circumstances and survival from childhood acute lymphoblastic leukaemia in West Germany.
Veröffentlichungen- Deutsches Kinderkrebsregister
Cancer epidemiology. Childhood cancer incidence patterns by race, sex and age for a report from the South African National Cancer Registry. Radiotherapy and subsequent thyroid cancer in German childhood cancer survivors: a nested case-control study. Radiation oncology London, England. Grabow D, P K. Pediatric germ cell tumors from to incidence rates, time trends, and survival.
Cancer spectrum and frequency among children with Noonan, Costello, and cardio-facio-cutaneous syndromes. British journal of cancer.
Risk of cancer incidence before the age of 15 years after exposure to ionising radiation from computed tomography: results from a German cohort study. Radiotherapy and subsequent thyroid cancer in German childhood cancer survivors. Registration of childhood cancer: Moving towards pan-European coverage?
Strategies to improve the quality of survival for childhood brain tumour survivors. Pediatric second primary malignancies after retinoblastoma treatment. Childhood cancer survivor cohorts in Europe. Parental occupational pesticide exposure and the risk of childhood leukemia in the offspring: findings from the childhood leukemia international consortium. Parental occupational paint exposure and risk of childhood leukemia in the offspring: findings from the Childhood Leukemia International Consortium. Rare malignant pediatric tumors registered in the German Childhood Cancer Registry Survival from childhood acute lymphoblastic leukaemia in West Germany: does socio-demographic background matter?
Local radiation dose and solid second malignant neoplasms after childhood cancer in Germany: a nested case-control study. Parental occupational exposure and risk of childhood central nervous system tumors: a pooled analysis of case-control studies from Germany, France, and the UK. Childhood leukaemia risks: from unexplained findings near nuclear installations to recommendations for future research. Maternal supplementation with folic acid and other vitamins and risk of leukemia in offspring: a Childhood Leukemia International Consortium study. Childhood and parental diagnostic radiological procedures and risk of childhood brain tumors.
Journal of neuro-oncology. Monatsschr Kinderh. Malignant ectomesenchymoma in children and adolescents: report from the Cooperative Weichteilsarkom Studiengruppe CWS.
Does socio-demographic background matter? Survival from paediatric acute lymphoblastic leukaemia in Germany. Survival and cure trends for European children, adolescents and young adults diagnosed with acute lymphoblastic leukemia from to Kaatsch P, Spix C. German Childhood Cancer Registry - Report Mainz: German Childhood Cancer Registry; The Childhood Leukemia International Consortium.
Michaelis J, Kaatsch P. Deutsches Kinderkrebsregister. Fetal growth and childhood acute lymphoblastic leukemia: findings from the childhood leukemia international consortium. Parental informed consent in pediatric cancer trials: a population-based survey in Germany. Childhood acute lymphoblastic leukaemia and birthweight: insights from a pooled analysis of case-control data from Germany, the United Kingdom and the United States. Effective childhood cancer treatment: the impact of large scale clinical trials in Germany and Austria.
Spix C. Spix C, Kaatsch P. Langzeitfolgen im Fokus. Searching for gene expression differences in primary fibroblasts between patients with one and two neoplasms in childhood. Pediatric hematology and oncology. Suspected infertility after treatment for leukemia and solid tumors in childhood and adolescence. Deutsches Arzteblatt international. Brecht I, Kaatsch P. Rare tumors in children and adolescents.
The differentiation of the strains is the basis of research into the epidemiology of infectious diseases. It is important to identify the sources of micro- epidemics and for the identification of clones with special properties, to spread an epidemic such as increased virulence or increased ability.
The initial genetic event which is marked the emergence of a MRSA strain, the acquisition of the mec element, which introduces the determinant of the mecA methicillin resistance in MRSA strain. Four types of mec elements or SCCmec have been identified so far. Jahrhunderts verzeichnet. Individual strains of MRSA are distinguished by their ability to rapidly spread.
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These strains are called epidemic MRSA strains and pose a serious risk to the patient is in the hospital, the rapid identification of epidemic MRSA can be a significant advantage in the fight against their spread and consequently this lead to a reduction in infection rates and ultimately mortality. Most epidemic strains occur in Europe and particularly in Germany. They are designated according to their multilocus sequence types by ST numbers. These strains occur only recently. It is very desirable to be able to distinguish between these new strains.
The method is labor intensive and time consuming. It comprises the cultivation of the organism, usually over night in a nutrient broth, embedding the washed cells in agarose, lysing of the cells in situ, digestion of the DNA in situ with a restriction enzyme, loading of a gel with the agar block, and subsequent separation of the fragments on the gel by PFGE. The whole procedure has been going without culture 2 to 5 days and this period is much too long for a much awaited in a hospital environment, identification of Staphylococcus aureus.
In addition, the method is so sophisticated that it will not be used in an ordinary hospital Tang et al, J.
This internationally recognized method is currently and J. Some strains of Staphylococcus aureus are only against one or two phages of the international standard sets sensitive, or can also completely insensitive to these phages be Zierdt CH et al J. In particular strains of Staphylococcus aureus which are sensitive only to one or two phages may be maintained on the basis of phage typing for identical incorrectly. Rapid and reliable differentiation of strains is important for the identification of sources of micro- epidemics, particularly in hospitals.
It is very important that you can identify clones with special properties, such as increased virulence or increased ability to spread rapidly epidemic. Previous attempts to identify a nucleic acid sequence which would be suitable for a clear and rapid differentiation of strains have failed. Some newer, also commercially operated, methods use the sequencing of DNA. However, this is tedious and time consuming. Until now represents the Spa Typing is not an alternative, which is based on the fact that a positive identification was not possible. Although Spa typing in the art as a method for the identification of Staphylococcus aureus is known, it has been but never used as a single proof of specific strains.
One older document teaches the contrary, that the sequences derived from this region would not be specific to a tribe. So far so is the spa Typing is not an alternative, which is based on the fact that a positive identification was not possible. Consequently, it would be desirable to know sequences, substances and methods to identify compared with the prior art said new strains of Staphylococcus aureus in a more differentiating and faster way.
Ideally, said sequences, substances and methods would make it possible to perform the analysis in a reliable manner in an average hospital environment. Furthermore, said sequences, substances and methods would not require cultivation of Staphylococcus aureus isolates for a long time. The present invention relates to a novel nucleic acid molecule selected from the group of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20 or a reverse complement thereof.
Table 1 lists both the numbers and names of the corresponding sequences as well as the number of the strain, for which the sequence is specific to. In connection with the present invention, nucleic acid molecules of the invention are used to uniquely determine the appropriate strain. Here, the terms nucleic acid molecule and oligonucleotide refer to both primers, probes Probes , sample and fragments of oligonucleotides, as well as other forms of nucleic acid molecules.
The terms nucleic acid molecule and oligonucleotide is further generic for Polydeoxribonucleotide containing 2-deoxy-Dribose and for polyribonucleotides containing D-ribose , or any other form of polynucleotide which is an N-glycoside of a purine or pyrimidine base, or modified purine base or pyrimidine is used.
The terms also include polyamides with a purine or pyrimidine base of which are also known as PNAs. Here are intended to include any length both nucleic acid molecule and oligonucleotide molecules, but in each case the included molecules very well have a defined length in turn.
The inventors have found that, in contrast to most other originating from Staphylococcus aureus Spa sequences which are disclosed in this invention, sequences of the invention can actually serve an unambiguous determination of certain strains of Staphylococcus aureus, however Table 1. Tabelle 1: Table A preferred embodiment of the invention relates to certain parts of the SEQ ID NO of the complete sequences: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20, respectively, or a reverse complement thereof, as shown in Table 2 below.
In one embodiment, the present invention relates to an oligonucleotide or a molecule comprising an oligonucleotide refers to the detection of one of the nucleic acid molecules of the invention according to the above description.
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NO: 21, 22, 23, 24, 25, 26, 27, 28, and 29, respectively, or the reverse complementary sequence thereof see also Table 2. The expert is here aware that the sequences of the oligonucleotides may vary slightly from those here. Likewise, one or a few nucleotides may be different. However, it is important that the specificity of the oligonucleotide is maintained and the melting temperature does not change significantly.
This means that when changing the sequence of the oligonucleotide, the melting temperature preferably remains unchanged. The skilled worker is also aware of this, that even modified nucleotides may account for a part or all of an oligonucleotide according to the invention. Capture Probes modifiziert. As apparent from the above section, the oligonucleotides are such as so-called on its 5 'end with an amino group to bind to surfaces in a preferred embodiment. Capture Probes modified. In one embodiment, the ingredients listed in Table 2 oligonucleotides are used together with a bound to the 5'-end mer poly T tail and either on glass slides, nylon or another solid support gespotted.
Here, the tail can also have between 2 and 50 nucleotides and consist of poly A, C, G, or T or a mixture thereof. Longer tails are possible but not ideal. In another embodiment, each chemically modified oligonucleotides are selected from the group consisting of a chemical modification for the immobilization of oligonucleotides, and a modification for the identification of the oligonucleotide. In a preferred embodiment of the invention, the oligonucleotide can in addition have still a chemical modification or a labeling group label group selected from the group comprising an amino linker, a fluorescent dye such as fluorescein or cyanine dye, biotin, digoxigenin, or others known to those skilled modifications.
The modification may serve as a binding site for a solid phase or serve to receive a label. Beispielsweise kann ein mit Biotin markiertes Oligonucleotid mit einem Enzym nachgewiesen werden, das seinerseits wiederum an Streptavidin gebunden ist. For example, a biotin labeled oligonucleotide can be detected with an enzyme, which in turn is attached to streptavidin.
Solch ein Enzym kann alkalische Phosphatase oder Peroxidase sein. Such an enzyme may be alkaline phosphatase or peroxidase.
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The reaction will thus result in a color change. Die Modifikation kann auch ein Linker wie beispielsweise ein Aminolinker sein. The modification can also be a linker such as an amino linker. However, the markers may also be a radioisotope such as I, 35 C, 32 P or 35 P.