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14. Evaluation of different target genes for the detection of Salmonella sp. by loop‐mediated isothermal amplification.

Author

Kreitlow, A., Becker, A., Schotte, U., Malorny, B., Plötz, M., and Abdulmawjood, A.

Subjects
GENE amplification, GENE targeting, SALMONELLA enterica, GENES, SINGLE nucleotide polymorphisms, SALMONELLA detection
Abstract

The loop‐mediated isothermal amplification (LAMP) technique was used to investigate six salmonella‐specific sequences for their suitability to serve as targets for the pathogen identification. Sequences selected for designing LAMP primers were genes invA, bcfD, phoP, siiA, gene62181533 and a region within the ttrRSBCA locus. Primers including single nucleotide polymorphisms were configured as degenerate primers. Specificity of the designed primer sets was determined by means of 46 salmonella and 32 other food‐ and waterborne bacterial reference species and strains. Primers targeting the ttrRSBCA locus showed 100 % inclusivity of target and exclusivity of other test species and strains. Other primer sets revealed deficiencies, especially regarding Salmonella enterica subsp. II–IV and Salmonella bongori. Additionally, primers targeting the siiA gene failed to detect S. enterica subsp. enterica serotypes Newport and Stanley, whereas bcfD primers did not amplify DNA of S. enterica subsp. enterica serotype Schleissheim. TtrRSBCA primers, providing short detection times and constant melting temperatures of amplification products, achieved best overall performance. [ABSTRACT FROM AUTHOR]

Published
2021
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20. Chlamydiaceae and chlamydia-like organisms in free-living small mammals in Europe and Afghanistan

Author

Stephan, S, Guerra, D, Pospischil, A, Hilbe, Monika; https://orcid.org/0000-0002-4133-2402, Weissenböck, H, Novotny, L, Greub, Gilbert; https://orcid.org/0000-0001-9529-3317, Croxatto, A, Teifke, J P, Ulrich, R G, Schlegel, M, Ruhl, S, Schotte, U, Binder, A, Sauer, S, Borel, Nicole; https://orcid.org/0000-0002-1556-9262, Stephan, S, Guerra, D, Pospischil, A, Hilbe, Monika; https://orcid.org/0000-0002-4133-2402, Weissenböck, H, Novotny, L, Greub, Gilbert; https://orcid.org/0000-0001-9529-3317, Croxatto, A, Teifke, J P, Ulrich, R G, Schlegel, M, Ruhl, S, Schotte, U, Binder, A, Sauer, S, and Borel, Nicole; https://orcid.org/0000-0002-1556-9262

Abstract

Few data are available on the occurrence of chlamydial infections in wild small mammals. Therefore, the present study aimed to investigate the significance of free-living small mammals as reservoirs or transmission hosts for chlamydiae. In total, 3,664 tissue samples originating from 911 animals were collected in Switzerland, Germany, Austria, the Czech Republic and Afghanistan. Samples included internal organs (n = 3,652) and feces (n = 12) from 679 rodents (order Rodentia) and 232 insectivores (order Eulipotyphla) and were tested by three different TaqMan real-time polymerase chain reactions (PCR) specific for members of the family Chlamydiaceae and selected Chlamydia-like organisms such as Parachlamydia spp. and Waddlia spp. Only one out of 911 (0.11%) animals exhibited a questionable positive result by Chlamydiaceae specific real-time PCR. Furthermore, five out of 911 animals (0.55%) were positive by specific real-time PCR for Parachlamydia spp. but could not be confirmed by Parachlamydia acanthamoebae secY qPCR. One out of 746 animals (0.13%) showed a positive result by real-time PCR for Waddlia chondrophila. This result was confirmed by Waddlia secY qPCR. This study represents the first detection of Chlamydia-like organisms in small wildlife in Switzerland. Considering previous negative results for Chlamydiaceae in wild ruminant species from Switzerland, these data suggest that wild small mammals are unlikely to be important carriers or transport hosts for Chamydiaceae and Chlamydia-like organisms.

Published
2014

27. Assessment of frequencies of lifestyle factors and polymorphisms of drug-metabolizing enzymes (NAT2, CYP2E1) in human hepatocellular carcinoma (HCC) patients in a department of surgical medicine--a pilot investigation

Author

Farker K, Hoffmann A, Scheele J, and Schotte U

Subjects
Male, medicine.medical_specialty, Pathology, Carcinoma, Hepatocellular, Alcohol Drinking, Arylamine N-Acetyltransferase, Pilot Projects, Gastroenterology, PstI, Risk Factors, Internal medicine, Genotype, medicine, Humans, Pharmacology (medical), Genotyping, Life Style, Pharmacology, Polymorphism, Genetic, biology, business.industry, Liver Neoplasms, Smoking, Heterozygote advantage, Cytochrome P-450 CYP2E1, Middle Aged, medicine.disease, Genotype frequency, Hepatocellular carcinoma, Etiology, biology.protein, Female, Restriction fragment length polymorphism, business
Abstract

The pathogenesis of human hepatocellular carcinoma (HCC) is a multistage process with the involvement of a multifactorial etiology. The role of drugs as risk factors has not been conclusively ascertained, but it appears that the use of oral contraceptives can be included. In the multifactorial etiology of human hepatocellular carcinoma (HCC ), an association and interaction between genetic polymorssphisms of xenobiotic metabolizing enzymes, lifestyle factors and cancer risk has been postulated. This pilot investigation examines the frequency of polymorphisms in selected genes (NAT2, CYP2E1) coding for xenobiotic metabolizing enzymes, and lifestyle habits (cigarette smoking, alcohol consumption) in 38 HCC patients. Genotyping of xenobiotic metabolizing enzymes was carried out using polymerase chain reaction restriction fragment length polymorphism methods and DNA extracted from peripheral blood cells. In addition, HCC patients were interviewed with regard to their cigarette smoking habits and alcohol consumption using a standardized questionnaire. The results ofthis pilot investigation showed that the majority of the HCC patients smoke and consume alcohol. We found no predominance of slow acetylators (45%) or rapid acetylators (55%). 70.6% of slow acetylators were smokers. 86.5% of all patients with homozygote PstI/RsaI genotype also carried the homozygote Dral genotype, whereas 10.8% of all subjects with heterozygote PstI/RsaI genotype also carried the heterozygote Dral genotype. These genotype frequencies remain to be confirmed in a larger ethnic group. Whether polymorphisms of xenobiotic metabolizing enzymes is an important risk factor in (cigarette smoking-/alcohol consumption) HCC or not is currently being investigated in a case-control study in the same ethnic group.

Published
2002

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