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NanoDrop (Thermo Fisher Scientific, Massachusetts, USA) and stored at −20 °C, to be used as templates in PCRs. We performed two multiplex PCRs to detect the eight genes encoding carbapenemases. The first multiplex PCR targeted three genes coding for class D
Zhong, X. Y., Holzgreve, W., Hahn, S.: Risk free simultaneous prenatal identification of fetal Rhesus D status and sex by multiplex real-time PCR using cell free fetal DNA in maternal plasma. Swiss. Med. Wkly, 2001, 131 , 70
. , Soubrier , M. P. , Liautard , J.P. : IS6501-anchored PCR for the detection and identification of Brucella species and strains . J Appl Bacteriol 81 , 154 – 160 ( 1996 ). 5
Környezetből származó mikrobiális nukleinsavminták vizsgálati lehetőségei
Examination possibilities of microbial nucleic acid samples derived from the environment
metagenomics: from microbiology to DNA sequencing technologies and bioinformatics. Front Genet. 2015; 6: 348. 8 Jeong J, Mun S, Oh Y, et al. A qRT-PCR method capable of
biochemical tests without adequate laboratory testing is not possible as pathogen-specific clinical symptoms are lacking. The introduction of molecular-based detection methods such as polymerase chain reaction (PCR) and real-time PCR assays that are sensitive
of North America and development of a differential PCR-RFLP assay to detect and differentiate infections between PCV-1 and PCV-2. J. Clin. Microbiol. 38, 2494--2503. Genetic characterization of type-2 porcine circovirus (PCV
The aim of this study was to determine the prevalence of Vibrio parahaemolyticus in shellfish samples harvested along the Slovenian coast. Shellfish samples of Mediterranean mussels (Mytilus galloprovincialis) were collected along the Slovenian coast at four locations (Seča, Piran, Strunjan and Debeli Rtič) between 2006 and 2008. Samples were examined and analysed for the presence of V. parahaemolyticus by conventional and molecular methods. The presence of Vibrio in the samples was examined by conventional methods on plate grown bacterial cells before and after enrichment in alkaline saline peptone water (ASPW). PCR methods were used for the detection of V. parahaemolyticus-specific toxR and tlh genes and of the virulence-associated tdh and trh genes. Out of 168 samples examined, 24 were positive for toxR and tlh genes by PCR from enrichment broth. Five out of 62 (8.1%), 4 out of 32 (12.5%) and 15 out of 74 (20.2%) samples were positive in 2006, 2007 and 2008, respectively. Colonies of V. parahaemolyticus were isolated from only one sample positive for V. parahaemolyticus by PCR.
During a five-year period (2000 to 2004) 74,342 pigs were tested by the intradermal tuberculin test in Croatia. Of them, 248 (0.33%) pigs were positive and 91 (0.12%) were found to be suspicious in 7 out of the 13 farms included in the study. Gross pathological changes characteristic of tuberculosis were observed in tuberculin-positive and/or suspicious swine. Mycobacterium was isolated from the lymph nodes of 183 out of 234 swine (78.2%). For better epidemiological understanding, isolates were typed by conventional methods, PCR and hybridisation. The results show that most of the isolates belonged to the Mycobacterium avium complex (175 isolates, 95.7%). Other isolates belonged to M. fortuitum (6 isolates, 3.3%), M. chelonae (1 isolate, 0.5%), and M. peregrinum (1 isolate, 0.5%). Isolated strains of the M. avium complex were identified as M. a. avium (37 isolates, 21.1%) and M. a. hominissuis (138 isolates, 78.9%).
This study aimed to test the efficacy of samplings for the detection of Haemophilus parasuis after metaphylactic treatment and subsequent challenge using an established model for Glässer’s disease. In this model, 36 piglets were equally assigned to a negative control, a positive control, and two trial groups receiving tulathromycin 7 or 4 days prior to challenge. The piglets of three groups were challenged intratracheally with H. parasuis serovar 5. As a result, four pigs in each challenged group died or had to be euthanised within 10 days post challenge. The remaining 15 pigs of these challenged groups survived until termination of the experiment (days 14–15). All pigs were necropsied and collective swabs of serosal surfaces were tested by bacterial culture and PCR. Samples of tarsal synovial fluid and joint capsule, cerebrospinal fluid (CSF) and brain swabs were tested by PCR. A total of 22 out of the 27 challenged animals had macroscopically detectable polyserositis and all of them tested positive in the collective swab samples. Haemophilus parasuis was more frequently detected in pigs that died within the first 10 days compared to those surviving until days 14–15 (P < 0.001), and those that succumbed within 10 days showed higher positivity rates in the brain and CSF. All pigs which were positive in the CSF had detectable meningitis. At days 14–15, joint samples from 5 of the remaining 15 pigs tested positive for H. parasuis. Four of these five animals did not show any macroscopic or histological lesions in the joints. In conclusion, collective swabs were the best sample material in acute cases, whereas samples from the joints gave the best results in chronic cases. In this challenge model it was not possible to prove the metaphylactic effect of tulathromycin administered 4 and 7 days prior to infection with H. parasuis.
reverse transcription-PCR to reveal cellular changes during stimuli that result in Herpes Simplex Virus type 1 reactivation from latency. J. Virol., 1998, 72 , 1252–1261. Lasner T. M
