scholarly journals Preliminary Study of Sars-Cov-2 Occurrence in Wastewater in the Czech Republic

2020 ◽  
Vol 17 (15) ◽  
pp. 5508 ◽  
Author(s):  
Hana Mlejnkova ◽  
Katerina Sovova ◽  
Petra Vasickova ◽  
Vera Ocenaskova ◽  
Lucie Jasikova ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Wastewater Treatment ◽  
Czech Republic ◽  
Wastewater Treatment Plants ◽  
Chain Reaction ◽  
The Czech Republic ◽  
Untreated Wastewater ◽  
Preliminary Study ◽  
Polymerase Chain

The virus SARS-CoV-2, which has caused the recent COVID-19 pandemic, may be present in the stools of COVID-19 patients. Therefore, we aimed to detect SARS-CoV-2 in wastewater for surveillance of SARS-CoV-2 in the population. Samples of untreated wastewater were collected from 33 wastewater treatment plants (WWTPs) of different sizes within the Czech Republic. SARS-CoV-2 RNA was concentrated from wastewater and viral RNA was determined using real-time reverse transcription polymerase chain reaction (RT-qPCR). SARS-CoV-2 RNA was detected in 11.6% of samples and more than 27.3% of WWTPs; in some of them, SARS-CoV-2 was detected repeatedly. Our preliminary results indicate that an epidemiology approach that focuses on the determination of SARS-CoV-2 in wastewater could be suitable for SARS-CoV-2 surveillance in the population.

scholarly journals PCR detection of Bartonella spp. in the dog

2014 ◽  
Vol 83 (2) ◽  
pp. 79-82 ◽  
Author(s):  
Jarmila Konvalinová ◽  
Vlasta Svobodová ◽  
Dobromila Molinková ◽  
Miroslav Svoboda
Keyword(s):  
Polymerase Chain Reaction ◽  
Czech Republic ◽  
Clinical Symptoms ◽  
Bartonella Henselae ◽  
Chain Reaction ◽  
The Czech Republic ◽  
Two Samples ◽  
Healthy Dogs ◽  
Polymerase Chain ◽  
First Time

Our study aimed at using PCR to identify the incidence ofBartonellaspp. in blood of dogs. Altogether 286 dogs of 92 breeds aged 3 month to 17 years were tested from October 2008 to December 2009. Healthy dogs as well as dogs with various clinical symptoms of disease were included in the group. Samples were tested by polymerase chain reaction (PCR) specific for the presence ofBartonellaspp. Following the DNA examination in 286 dogs by PCR and subsequent sequencing, two samples were identified asBartonella henselae(0.7%). Other species ofBartonellawere not found. It was the first time in the Czech Republic when incidence ofBartonellaspp. was determined in dogs.

scholarly journals Detection of selected antibiotic resistance genes using multiplex PCR assay in mastitis pathogens in the Czech Republic

2017 ◽  
Vol 86 (2) ◽  
pp. 167-174 ◽  
Author(s):  
Vladimir Pyatov ◽  
Irena Vrtková ◽  
Aleš Knoll
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Czech Republic ◽  
Resistance Genes ◽  
Multiplex Polymerase Chain Reaction ◽  
Antibiotic Resistance Genes ◽  
Chain Reaction ◽  
The Czech Republic ◽  
Polymerase Chain ◽  
Mastitis Pathogens

The aim of this research was to develop multiplex polymerase chain reaction assays for the detection of aminoglycoside (strA, strB), sulphonamide (sulI, sulII), tetracycline (tetA, tetB, tetK, tetM, tetO), macrolide and lincosamide (msrA, ermA, ermB, ermC, mefA/E) genes of resistance in mastitis pathogens (Escherichia coli, Staphylococcus aureus, Streptococcus uberis, Streptococcus agalactiae and Streptococcus dysgalactiae). Applying the established assays, we investigated the distribution of antibiotic resistance genes in the above mentioned species isolated from milk samples in the Czech Republic. Each assay consisted of seven pairs of primers. Six of them amplified fragments of antibiotic resistance genes and one pair a fragment of a species specific gene. Polymerase chain reaction conditions were optimized to amplify seven gene fragments simultaneously in one reaction. In total, 249 isolates were used, among which 111 were positive for E. coli, 52 for S. aureus and 86 for Streptococcus spp. The majority (60.2%) of bacteria carried at least one antibiotic resistance gene and 44.6% were multidrug-resistant. The designed multiplex polymerase chain reaction assays may be applied as diagnostic method to replace or complement standard techniques of antibiotic susceptibility testing in the mentioned pathogens.

scholarly journals Distribution of Two Formae Speciales of Polymyxa Graminis in the Czech Republic

2017 ◽  
Vol 48 (2) ◽  
pp. 54-62
Author(s):  
L. Grimová ◽  
L. Winkowska ◽  
B. Špuláková ◽  
P. Růžičková ◽  
P. Ryšánek
Keyword(s):  
Polymerase Chain Reaction ◽  
Czech Republic ◽  
Soil Samples ◽  
Chain Reaction ◽  
The Czech Republic ◽  
Polymyxa Graminis ◽  
Formae Speciales ◽  
Crop Fields ◽  
Polymerase Chain ◽  
Mixed Populations

Abstract It has been shown that two formae speciales of P. graminis, namely f. sp. temperata (ribotype Pg-I) and f. sp. tepida (ribotype Pg-II), are widely distributed throughout temperate areas of Europe. In this study, the presence of both forms of the temperate Polymyxa spp. was identified in soil samples from different locations of the Czech Republic during a survey performed in 2012 and 2013. Based on polymerase chain reaction results, of the total 58 tested samples, 67.2% contained at least one monitored forma specialis. Specifically, P. graminis f. sp. temperata was detected in 48.3% of soil samples, while P. graminis f. sp. tepida was detected in 44.8% of samples. Mixed populations were found in 25.9% of the tested areas. This plasmodiophorid was confirmed not only in crop fields but also in meadows and forests in all explored regions. Our results extend the knowledge on the distribution of both ribotypes of P. graminis and provide the first evidence of f. sp. tepida within the Czech Republic.

scholarly journals Diagnostic efficacy of molecular assays for the viral haemorrhagic septicaemia virus isolates from the Czech Republic

2017 ◽  
Vol 86 (3) ◽  
pp. 207-212 ◽  
Author(s):  
Ľubomír Pojezdal ◽  
Dagmar Pokorová ◽  
Stanislava Reschová ◽  
Miroslava Palíková ◽  
Monika Vícenová ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Czech Republic ◽  
Real Time ◽  
Reverse Transcription ◽  
Chain Reaction ◽  
The Czech Republic ◽  
Haemorrhagic Septicaemia ◽  
Viral Haemorrhagic Septicaemia Virus ◽  
Polymerase Chain ◽  
The One

The diagnostic properties of the one-step real-time reverse-transcription polymerase chain reaction assay for viral haemorrhagic septicaemia virus detection were compared to methods currently in use in the Czech Republic, namely, virus isolation using the cell culture and conventional reverse-transcription polymerase chain reaction followed by the nested polymerase chain reaction. The assays were tested on a panel of 25 archived viral haemorrhagic septicaemia isolates and 8 archived infectious haematopoietic necrosis isolates obtained from monitoring and/or outbreaks of the diseases among farmed salmonids in the Czech Republic. The ability to detect the presence of the virus in the tissues of fish was tested on additional 32 field samples collected from the rainbow trout (Oncorhynchus mykiss), brown trout (Salmo trutta) and brook trout (Salvelinus fontinalis). The real-time assay showed the highest analytic sensitivity by detecting the presence of viral nucleic acid in samples with 10-7 dilution, whereas the sensitivity of the conventional polymerase chain reaction peaked at 10-5. Diagnostic specificity of both molecular assays was confirmed by absence of cross-reactivity with the infectious haematopoietic necrosis virus isolates. This, along with consistent results in the detection of the virus in the fish tissues, confirms that the one-step real-time reverse-transcription polymerase chain reaction is currently an optimal stand-alone diagnostic method for the detection of the viral haemorrhagic septicaemia virus.

Identification of Bovine-Specific DNA in Feedstuffs

2001 ◽  
Vol 64 (1) ◽  
pp. 117-119 ◽  
Author(s):  
PAVEL KRČMÁŘ ◽  
EVA RENČOVÁ
Keyword(s):  
Polymerase Chain Reaction ◽  
Mitochondrial Dna ◽  
Czech Republic ◽  
Dna Sequences ◽  
Fish Meal ◽  
Spongiform Encephalopathy ◽  
Vertebrate Species ◽  
Chain Reaction ◽  
The Czech Republic ◽  
Polymerase Chain

Considering the menace of transmission of bovine spongiform encephalopathy, feed components intended for cattle nutrition must be checked for the presence of bovine-derived materials. We have been using a method based on polymerase chain reaction for the identification of bovine-specific mitochondrial DNA sequences for this purpose. The specificity of the primers for polymerase chain reaction has been tested using samples of DNA of other vertebrate species, which may also be present in rendering plant products. The method allows the detection in concentrate mixtures of 0.125% of bovine-derived material. Bovine DNA at concentrations corresponding to less than 0.5% of bovine-derived material was detected in 3 of the 30 samples of concentrate mixtures collected from distributors' stores all over the Czech Republic. All 44 samples of fish meal collected from the same sources were free of bovine-derived material.

scholarly journals Assessment of Aggregatibacter actinomycetemcomitans and Prevotella intermedia Counts around Healthy Implants, Diseased Implants and Sound Teeth: A Preliminary Study

2017 ◽  
Vol 9 (1) ◽  
pp. 18-25 ◽  
Author(s):  
Siamak Yaghobee ◽  
Afshin Khorsand ◽  
Nojan Jahedmanesh ◽  
Mahdi Kadkhodazadeh
Keyword(s):  
Polymerase Chain Reaction ◽  
Gingival Crevicular Fluid ◽  
Aggregatibacter Actinomycetemcomitans ◽  
Prevotella Intermedia ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Standard Curve ◽  
Preliminary Study ◽  
Polymerase Chain ◽  
Crevicular Fluid

Background. This study aimed to assess Aggregatibacter actinomycetemcomitans (Aa) and Prevotella intermedia (PI) counts in gingival crevicular fluid (GCF) around healthy implants, diseased implants and sound teeth. Methods. Eight patients (four males and four females), who had healthy implants, implants with peri-implantitis and sound teeth, were selected. Samples (GCF) were analyzed using real-time polymerase chain reaction (RT-PCR). The above-mentioned bacteria were detected and counted. Data analysis in RT-PCR was carried out based on the standard curve using Prism software to compare Pi and Aa counts between the three areas (GCF around sound teeth, healthy implants and implants with peri-implantitis). Results. Pi counts were significantly higher in GCF around implants with peri-implantitis (8 implants) than around healthy implants (8 implants) (P<0.001) and sound teeth (8) (P=0.012). No significant differences were found in Pi counts in GCF around healthy implants and sound teeth (P=0.063). Aa counts in GCF around implants with peri-implantitis were significantly higher than those around healthy implants (P=0.002) and sound teeth (P=0.024). No significant differences were noted in Aa counts in GCF around healthy implants and sound teeth (P=0.57). Conclusion. Aa and Pi counts in GCF around diseased implants were higher than around healthy implants and sound teeth. Also, Aa counts were significantly higher than Pi counts.

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