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combined PCR-RFLP method. Dis. Aquat. Org. 44, 35--39. Identification of fish parasitic Myxo-bolus (Myxosporea) species using a combined PCR-RFLP method. Dis. Aquat. Org

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., Kwok-Yung, Y., Wing-Cheong, Y.: Detection of KatG Ser315Thr substitution in respiratory specimens from patients with isoniazid-resistant Mycobacterium tuberculosis using PCR-RFLP. J. Med. Microbiol. 52 , 999–1003 (2003

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Acta Alimentaria
Authors: J. Krulj, N. Ćurčıć, A. Bočarov Stančıć, J. Kojıć, L. Pezo, L. Peıć Tukuljac, and M. Bodroža Solarov

sequences for the description and identification of the species using a multilocus approach. PCR-restriction fragment length polymorphism (PCR-RFLP) is a simple, cost effective and quick tool for rapid detection of specific differences in DNA sequences of

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Acta Veterinaria Hungarica
Authors: Ljiljana Kuruca, Aleksandra Uzelac, Ivana Klun, Vesna Lalošević, and Olgica Djurković-Djaković

. , Dubey , J. P. and Su , C. ( 2014 ): Geographical patterns of Toxoplasma gondii genetic diversity revealed by multilocus PCR-RFLP genotyping . Parasitol. 141 , 453 – 461

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Acta Veterinaria Hungarica
Authors: Zsuzsanna Varga, Boglárka Sellyei, Petra Paulus, Melitta Papp, Kálmán Molnár, and Csaba Székely

The objective of this study was to survey the incidence of Flavobacterium columnare in wild and cultured freshwater fish species in Hungary. This bacterium usually causes disease in waters of more than 25 °C temperature. However, with the introduction of intensive fish farming systems, infected fish exposed to stress develop disease signs also at lower temperatures; in addition, the temperature of natural waters rises to the critical level due to global warming. Twenty-five isolates from wild and cultured freshwater fishes were identified as F. columnare by specific PCR, although both the fragment lengths and the results of PCRRFLP genotyping with BsuRI (HaeIII) and RsaI restriction enzymes raised doubts regarding this species classification. Sequencing of the 16S ribosomal RNA gene revealed that 23 isolates belonged to the species F. johnsoniae and two represented Chryseobacterium spp. The isolates were found to have high-level multidrug resistance: all were resistant to ampicillin and polymyxin B, the 23 F. johnsoniae strains to cotrimoxazole, 88% of them to gentamicin, and 72% to chloramphenicol. The majority of the 25 isolates were sensitive to erythromycin (88%), furazolidone (76%), and florfenicol (68%).

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matching. All isolates showing unique ERIC-PCR patterns were subjected to species identification. Species identification Aeromonas isolates were subjected to a lab-on-a-chip-based PCR-RFLP of the rpoD gene fragment for species identification ( Puah et al

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JPC - Journal of Planar Chromatography - Modern TLC
Authors: Mitja Križman, Jernej Jakše, Mirko Prošek, Dea Baričevič, and Branka Javornik

Agarose gel electrophoresis is a basic separation tool used in molecular biology, mostly for qualitative DNA analysis. There are constraints limiting its use in quantitative analysis, namely low repeatability and a narrow linear range. However, by using an internal standard or internal normalization, repeatability and linear range could be significantly improved. In the work discussed in this paper it was shown that an approximately fivefold improvement in repeatability and an over threefold wider linear range could be achieved by applying internal normalization. Using the proposed approach, genetic markers, for example RAPD and PCR-RFLP, or even microsatellite markers, could be conveniently quantitatively assessed using agarose gel electrophoresis.

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Acta Veterinaria Hungarica
Authors: Bernadett Khayer, Zsuzsanna Rónai, Enikő Wehmann, and Tibor Magyar

Four urease-negative Bordetella bronchiseptica isolates originating from pigs were examined by phenotypic and molecular methods. The phenotypic properties of the isolates were in harmony with the data of the literature, except for the lack of urease activity in conventional tube test, API 20 NE and Diatabs™ assays. Using genotypic methods, the urease-negative isolates did not differ from the urease-positive reference strain. They were positive in species-specific and ureC PCR, and all strains showed uniform bands in PCR-RFLP studies of flaA genes. The reason for the lack of urease activity, a characteristic considered speciesspecific for B. bronchiseptica, needs to be studied further. The finding underlines the significance of genotyping when the phenotypic identification of B. bronchiseptica seems questionable.

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Several phenotypic and genotypic methods are currently used to identify subtypes of Campylobacter jejuni isolates. Of the phenotypic methods one, i.e. serotyping based on the heat stable antigen, is often hindered by the relatively large number of un-typable (NT) strains. Little is known, however, about the heterogeneity of the group formed by these NT strains. Therefore we serotyped 92 Hungarian, non-outbreak C. jejuni isolates and subjected the 28 NT strains to molecular analysis using PCR-RFLP of the flaAgene and to pulsed-field gel electrophoresis. With both methods the NT strains were classified into several molecular types (17 and 25, respectively), while the number of subgroups based on the results of the two techniques combined was twenty- six. These results indicate that the NT group of strains is extremely heterogeneous in Hungary, and the epidemiological connection between two NT isolates cannot be established or excluded without the use of molecular typing techniques.

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A colorectalis rákok a második leggyakoribb halálokként szerepelnek a rosszindulatú betegségek között. A fej-nyak táji daganatok halálozása Magyarországon 265%-kal emelkedett az utóbbi 30 évben. Nem lehet eléggé hangsúlyozni e két daganatcsoport közegészségügyi jelentőségét. A colorectalis daganatok esetén a GSTM1, GSTT1 enzimek, valamint a p53 72-es kodonjának, fej-nyak táji tumorok esetén az XRCC1 Arg194Trp és Arg399Gln polimorfizmusainak hatását elemeztük. Intraoperatíve eltávolított daganatos és megfelelően illesztett daganatmentes mintákat válogattunk. A formalinban fixált mintákat deparaffinizáltuk és proteináz-K-emésztésnek vetettük alá. A genotipizálást PCR, illetve a fej-nyak táji tumorok esetén PCR-RFLP módszerrel végeztük. A vizsgált allélek gyakoriságában nem volt különbség a daganatos és a kontrollcsoport között. Túlélés tekintetében szignifikáns különbséget találtunk a GSTM1 és a p53 allélek között Dukes B stádiumú colorectalis daganatok esetén és az XRCC1 194 allélek között III-as stádiumú fej-nyak táji tumorokban. A fenti típusú genetikai különbségek szisztematikus vizsgálata a jövőben hozzájárulhat az egyéni rizikóbecslés és az individualizált kezelések megalapozásához.

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