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The aim of these studies was the clonal analysis of 48 S. aureus strains, isolated from milk samples from 48 cows with clinical and subclinical mastitis. Specific markers were detected using PCR: gap gene as the genus marker for Staphylococcus spp. and nuc gene as the species marker for S. aureus. Clonal typing of S. aureus isolates was carried out using RAPD-PCR with two primers simultaneously. Clonal relatedness was determined on the basis of the number and molecular mass of RAPD amplicons and analyzed using the unweighted pair group method with arithmetic mean (UPGMA) and Jaccard’s similarity coefficient. On the basis of RAPD-PCR and clonal analysis employing the UPGMA method with Jaccard coefficient it was determined that all of the analyzed S. aureus strains were identical on the genome level. This indicates that only one bacterial clonal type was responsible for mastitis caused by S. aureus in the analyzed herd. This denotes that the virulence of S. aureus isolated from mastitis cases is very high, and that the elimination of this strain is very difficult. The research showed the usefulness of RAPD-PCR in interspecies S. aureus strains typization and the prospect of employing it in routine epidemiological diagnostics of mastitis in cows. The contribution of S. aureus strains representing one clonal type in the etiology of mastitis in cows from one herd was also confirmed.
Campylobacter, especially C. jejuni, is now recognized as a major cause of bacterial enteritis in people in most developed countries. The diversity within C. jejuni species may be described by both phenotypic and genotypic methods. In recent years, molecular methods, such as the restriction of fragment length polymorphism (RFLP), fla typing or random amplified polymorphic DNA (RAPD), have been proposed as alternative tests to phenotypic methods for typing of Campylobacter. This study was carried out to investigate the genetic variation among isolates C. jejuni by RAPD-PCR. The analysis of 19 isolates of C. jejuni performed with two primers (1254 and OPA-11) generated multiple amplification products, with sizes between 300 and 2000 bp. The UPGMA analysis of the fingerprints classified these strains to 10 clonal groups. The discriminatory index D=0.73 indicates a high differentiation ability of the method. Such data are valuable in epidemiological studies and for investigating the distribution of genotypes in different environments.
Despite the permanent development of laboratory techniques, various types of adulterations are still a problem in the food industry. An important group among different frauds is adulterations connected with meat species authenticity. Uncovering of adulterated meat products is important inter alia for allergic individuals, for those who can’t intake certain species because of religious beliefs, and for maintaining fair-trade. More subtle techniques used for the mentioned adulterations generate a need for the elaboration of better analytical methods to provide effective control of meat and meat products. The aim of this review is to present currently used laboratory methods applied for meat species identification and detection of adulterations in the declared composition of meat products. The first group of the described methods enables identification and analysis of proteins and the second presented group contains techniques of DNA analysis. Apart from their short characteristics, some disadvantages and potential problems found during work with certain methods are described.
Dietetic and nutritional value of fish and shellfish products have made them very attractive food products for consumers. This results in both the increase of seafood consumption per capita and more diverse offers of finfish/shellfish species on the market. Nevertheless, while pursuing profits some of the producers/importers may replace valuable, more costly raw materials with cheaper substitutes. In order to efficiently monitor what is in the product and not to allow such practices to take place it is necessary to apply reliable, fast methods as to verify their contents. Until recently the methods used to test the products’ components were based mostly on electrophoretic, chromatographic or the immunological tests. At presents to detect food adulteration the PCR technique and its variety based on species polymorphy (multiplex PCR, PCR-RFLP, PCR-RAPD, PCR-AFLP, PCR-FIN or PCR-SSCP) are used. The latest achievements in identification and differentiation between the species are real-time PCR and microarray DNA. Discussions on how to improve the methods of identification/differentiation and on the possible application of such methods in routine quality control of seafood by the appropriate authorities are in progress.
Opracowano wiele testów wykorzystujących cechy morfologiczne, biochemiczne i fizjologiczne bakterii w celu ich identyfikacji i klasyfikacji. W artykule zaprezentowano krótki opis wybranych metod genetycznych stosowanych w identyfikacji, klasyfikacji i różnicowaniu mikroorganizmów: analizę polimorfizmu fragmentów po trawieniu enzymami restrykcyjnymi, elektroforezę w zmiennym polu elektrycznym, elektroforezę w gradiencie czynnika denaturującego, analizę polimorfizmu fragmentów pojedynczoniciowego DNA, amplifikację techniką PCR losowych fragmentów DNA. Opisane metody zwiększają możliwości analityczne ograniczone ilością komercyjnie dostępnych zestawów diagnostycznych, a tym samym pozwalają na rzetelną i szybką identyfikację mikroorganizmów.
Diagnostics of Mycoplasma synoviae (MS) is generally based on serological, culturing and molecular methods. Rapid diagnosis and identification of infections is very important in the poultry industry. The use of PCR or real-time PCR makes it possible to shorten the time for obtaining results from research and the effective detection of genetic material of MS. There are many variations of the PCR method, one of them is the LAMP method, which is rapid, very sensitive and does not require specialist equipment. This article reviews the molecular methods used for the diagnosis of Mycoplasma synoviae infection in poultry.
The inflorescences of yellow everlasting collected from twenty two natural sites in the middle part of the Bug river valley and two in the Puszcza Knyszyńska forest were compared in respect to the content and composition of flavonoids and phenolic acids. Genetic distances among populations, evaluated by RAPD markers, fluctuated from 0.0968 to 0.4855. There were no dependences between genetic relationships and geographical distributions of populations, as well as between their genetic relationships and chemical compositions. The total content of flavonoids ranged from 0.15 to 0.70% and phenolic acids from 0.45 to 1.52%. As a result of the HPLC analysis no flavonoid compounds were identified, with isosalipurposide as a dominant constituent. The chlorogenic, caffeic, ferulic and rosmarinic acids were also detected.
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