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The influence of LDHA gene polymorphism on relative level of its expression in racing pigeons

100%

Jedrzejczak-Silicka M., Yu Y.-H., Cheng Y.-H., Dybus A.

Acta Scientiarum Polonorum. Zootechnica

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2018

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tom 17

|

nr 3

2

Detection of porcine endogenous retrovirus in xenotransplantation

100%

Gola J., Mazurek U.

Reproductive Biology

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2014

|

tom 14

|

nr 1

3

Tracking chromatin states using controlled DNase I treatment and real-time PCR

88%

Martins R. P., Platts A. E., Krawetz S. A.

Cellular and Molecular Biology Letters

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2007

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tom 12

|

nr 4

545-555

A novel approach to DNase I-sensitivity analysis was applied to examining genes of the spermatogenic pathway, reflective of the substantial morphological and genomic changes that occur during this program of differentiation. A new real-time PCR-based strategy that considers the nuances of response to nuclease treatment was used to assess the nuclease susceptibility through differentiation. Data analysis was automated with the K-Lab PCR algorithm, facilitating the rapid analysis of multiple samples while eliminating the subjectivity usually associated with Ct analyses. The utility of this assay and analytical paradigm as applied to nuclease-sensitivity mapping is presented.

4

Application of SYBR Green I and TaqMan probe-based real-time PCRs for the identification of Listeria spp. and Listeria monocytogenes

88%

Kedrak-Jablonska A., Budniak S., Szczawinska A., Reksa M., Krupa M., Szulowski K.

Bulletin of the Veterinary Institute in Puławy

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2015

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tom 59

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nr 4

The aim of the study was the application and comparison of real-time PCR methods based on the fluorescence of SYBR Green I intercalating dye and TaqMan probes for the detection of the 23S rDNA gene of Listeria spp. and the hlyA gene of Listeria monocytogenes. Five strains of L. monocytogenes and single strains of each of the species: L. ivanovii, L. innocua, L. grayi, L. welshimeri, and L. seeligeri were used for the experiments. Additionally, five strains of other species of bacteria were used for evaluation of the specificity of the tests. QuantiTect SYBR Green PCR and QuantiTect Probe PCR kits were selected for the study. In the first stage of the study, SYBR Green I real-time PCRs were performed under several methods, the first one allowing detection of the 23S rDNA gene and the remainder based on the amplification of the hlyA gene. In the next part, three varied in method TaqMan probe-based real-time PCRs allowing confirmation of strains belonging to Listeria spp. and L. monocytogenes were conducted. The observation of amplification curves in real-time PCR methods enabled the detection of both genes, and these methods demonstrated a significant sensitivity and high specificity. A high regression coefficient of 0.99 was found for all reactions. Specific amplification products were obtained for the 23S rDNA and hlyA genes, which confirmed the tested strains as Listeria spp. and L. monocytogenes respectively. Isolates of other microbial species did not yield real-time PCR products.

5

Validating allergen coding genes (Cor a 1, Cor a 8, Cor a 14) as target sequences for hazelnut detection via real-time PCR

75%

Recupero M., Garino C., D'Andrea M., Locatelli M., Cereti E., Coisson J. D., Arlorio M.

Polish Journal of Food and Nutrition Sciences

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2011

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tom 61

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nr Suppl.1

6

Construction of new GFP-tagged fusants for Trichoderma harzianum with enhanced biocontrol activity

75%

Kowsari M., Motallebi M., Zamani R. M.

Journal of Plant Protection Research

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2014

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tom 54

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nr 2

Trichoderma is one of the most exploited biocontrol agents for the management of plant diseases. In biocontrol ecology, the critical factors are detection, and the monitoring and recovery of specific biocontrol agents either naturally present or deliberately released into the environment. Protoplast fusion is an appropriate tool for the improvement of biocontrol Trichoderma strains. Protoplast isolation from Trichoderma harzianum was achieved using 24 h culture age, 6.6 mg/ml Novazym L 1412 at 30°C which resulted the maximum protoplast yield of 5 × 108/ml. The self-fused protoplasts were regenerated and 12 fusants were selected based on their growth rate on 2% colloidal chitin medium. Next, a comparison was done for chitinase and antagonistic activity. Transcriptomic analysis based on quantitative real-time RT-PCR, demonstrated that T8-05 fusant expressed 1.5 fold of chit42 transcript as compared with the parental line. This fusant with 7.02±0.15U chitinase activity showed a higher growth inhibition rate (100%) than the parent strain, against Rhizoctonia solani. To obtain a genetically marked fusant that can be used as a biomonitor, the fusant was cotransformed with the gfp and amdS genes. The morphology and viability of selected cotransformant (FT8-7MK-05-2) was similar to the parent. Green fluorescing conidia were observed within the first 2 days of incubation in the soil, and this was followed by the formation of chlamydopores after 60 days. The colonisation of the gfp-tagged fusant was also monitored visually on R. solani sclerotia by scanning electron microscopy. Production of gfp-tagged fusant of Trichoderma spp. provides a potentially useful tool for monitoring hyphal growth patterns and the population of biocontrol agent isolates introduced into environmental systems.

7

Quantitative estimation of porcine endogenous retrovirus release from PK15 cells

75%

Kimsa M., Strzalka-Mrozik B., Kimsa M., Adamska J., Gola J., Lopata K., Mazurek U.

Polish Journal of Microbiology

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2012

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tom 61

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nr 3

The present study focuses on the assessment of porcine endogenous retrovirus (PERV) release from PK15 cells in a time dependent manner. the highest amount of PERV A RNA was detected in PK15 cells after 16 hours of culture. !e highest amount of PERV B RNA was detected in PK15 cells after 20 hours. !e highest amount of both subtypes RNAs was detected in culture medium after 32 hours of culture. the peaks of PERV reverse transcriptase (RT) activity were detected after 28 h of culture in PK15 cells and after 32 hours in the culture medium. the monitoring of PERV release from PK15 cell line may be useful for the evaluation of PERV replication.

8

Incidence of adenoviral DNAemia in Polish adults undergoing allogeneic haematopoietic stem cell transplantation

75%

Rynas S., Dzieciatkowski T., Przybylski M., Basak G. W., Rusicka P., Tomaszewska A., Halaburda K., Jedrzejczak W. W., Mlynarczyk G.

Archivum Immunologiae et Therapiae Experimentalis

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2015

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tom 63

|

nr 1

9

Real-time PCR and agar plating method to predict Fusarium verticillioides and fumonisin B1 content in Nigerian maize

75%

Adejumo T. O., Hettwer U., Nutz S., Karlovsky P.

Journal of Plant Protection Research

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2009

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tom 49

|

nr 4

399-404

Eighty maize grain samples collected in Nigeria were investigated for fumonisin B1 (FB1) content and Fusarium verticillioides colonization. F. verticillioides DNA was quantified by species-specific real-time PCR and living propagules of the fungus were counted by agar-plating method. FB1 was detected in 55 (68.7%) of the total samples (mean: 98.5 μg/kg, range: 10 to 714 μg/kg) at 10 μg/kg detection limit. The mean amount of F. verticillioides DNA determined by real-time PCR was 49.7 μg/kg (range: 10-126.7 μg/kg), while agar plate method showed the presence of F. verticillioides in 45 samples (mean incidence: 21.0%, range: 6.7-60.0%). There was correlation ties between F. verticillioides DNA by real time PCR and fungal colonization by agar plate method (R = 0.71, p = 00001 at 95% confidence level), and means of FB1 and F. verticillioides DNA in the yellow and white maize were significantly different. Despite the high consumption of maize in Nigeria, the amount of FB1 ingested by consumers appears to be low. The estimated daily intake of fumonisins was 0.21 μg/kg body weight per day.

10

False negative results in high viremia parvovirus B19-samples tested with real-time PCR

75%

Grabarczyk P., Kalinska A., Sulkowska E., Brojer E.

Polish Journal of Microbiology

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2010

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tom 59

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nr 2

129-132

Extremly high viremia is observed during some viruses infection, especialy in immunocompromised patients. False negative results of Parvovirus B19 DNA tests performed with real-time PCR in high viremic samples are reported. The way of fluorescence diagrams analysis and algorithm of positive result confirmation to exclude such phenomenon are proposed.

11

Co-infections with cytomegalovirus and human herpesvirus type 7 in adult Polish allogeneic haematopoietic stem cell transplant recipients

75%

Tomaszewska A., Krysko A., Dzieciatkowski T., Przybylski M., Basak G. W., Halabura K., Piekarska K., Sulowska A., Nasilowska-Adamska B., Mlynarczyk G., Jedrzejczak W. W., Marianska B.

Archivum Immunologiae et Therapiae Experimentalis

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2014

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tom 62

|

nr 1

12

Evaluation of methods for Erwinia amylovora detection

75%

Kaluzna M., Pulawska J., Mikicinski A.

Journal of Horticultural Research

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2013

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tom 21

|

nr 2

13

Cloning and expression of apyrase gene from Ancylostoma caninum in Escherichia coli

75%

Qiao Y., Pengsakul T.

Acta Parasitologica

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2015

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tom 60

|

nr 1

14

Reference genes for gene expression studies on non-small cell lung cancer

75%

Gresner P., Gromadzinska J., Wasowicz W.

Acta Biochimica Polonica

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2009

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tom 56

|

nr 2

307-316

Study Objective: The aim of this study was to test a panel of 6 reference genes in order to identify and validate the most suitable reference genes for expression studies in paired healthy and non-small cell lung cancer tissues. Method: Quantitative real-time PCR followed by the NormFinder- and geNorm-based analysis was employed. The study involved 21 non-small cell lung cancer patients. Results: The analysis of experimental data revealed HPRT1 as the most stable gene followed by RPLP0 and ESD. In contrast, GAPDH was found to be the least stable gene. HPRT1 together with ESD was revealed as the pair of genes introducing the least systematic error into data normalization. Validation by bootstrap random sampling technique and by normalizing exemplary gene expression data confirmed the results. Conclusion: Although HPRT1 and ESD may by recommended for data normalization in gene expression studies on non-small cell lung cancer, the suitability of selected reference genes must be unconditionally validated prior to each study.

15

New primers for fast detection of Giardia duodenalis assemblages A and B using real-time PCR

75%

Solarczyk P., Wojtkowiak-Giera A., Holysz M., Slotkowicz-Kowalska A., Jagodzinski P. P., Stojecki K., Rocka A., Majewska A. C., Skrzypczak L.

Acta Protozoologica

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2018

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tom 57

|

nr 1

16

New PCR test for detection on Candida glabrata based on the molecular target chosen by RAPD technique

75%

Olchawa A., Krawczyk B., Brillowska-Dabrowska A.

Polish Journal of Microbiology

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2013

|

tom 62

|

nr 1

Rapid, reliable diagnosis is a necessary condition for the successful treatment of infections. Such diagnostic assays are continually being developed. The paper presents a method for selecting the molecular target for PCR-based diagnostics based on the comparison of RAPD patterns. A sequence encoding Candida glabrata CBS138 hypothetical protein was selected. The limit of detection for PCR and real-time PCR reactions with DNA extracted from blood samples spiked with Candida glabrata was estimated at 1 CFU/ml. The application of the assays developed in this study would thus seem to be promising as a complementary method in the diagnostics of C. glabrata infections.

17

Detection and differentiation of Newcastle disease virus and influenza virus by using duplex real-time PCR

75%

Nidzworski D., Wasilewska E., Smietanka K., Szewczyk B., Minta Z.

Acta Biochimica Polonica

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2013

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tom 60

|

nr 3

Newcastle disease virus (NDV), member of the Paramyxoviridae family and avian influenza virus (AIV), member of the Orthomyxoviridae family, are two main avian pathogens causing serious economic problems in poultry health. Both are enveloped, single-stranded, negative-sense RNA viruses and cause similar symptoms, ranging from sub-clinical infections to severe diseases, including decrease in egg production, acute respiratory syndrome, and high mortality. Similar symptoms hinder the differentiation of infection with the two viruses by standard veterinary procedures like clinical examination or necropsy. To overcome this problem, we have developed a new duplex real-time PCR assay for the detection and differentiation of these two viruses. Eighteen NDV strains, fourteen AIV strains, and twelve other (negative control) strains viruses were isolated from allantoic fluids of specific pathogen-free (SPF), embryonated eggs. Four-weeks-old SPF chickens were co-infected with both viruses (NDV - LaSota and AIV - H7N1). Swabs from cloaca and trachea were collected and examined. The results obtained in this study show that by using duplex real-time PCR, it was possible to detect and distinguish both viruses within less than three hours and with high sensitivity, even in case a bird was co-infected. Additionally, the results show the applicability of the real-time PCR assay in laboratory practice for the identification and differentiation of Newcastle disease and influenza A viruses in birds.

18

In house validation of real-time PCR method for detection of Clostridium botulinum in food and feed matrixes

75%

Grenda T., Kwiatek K.

Bulletin of the Veterinary Institute in Puławy

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2010

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tom 54

|

nr 4

For validation purposes, characteristic parameters for quantitative detection were estimated according to the PN-EN ISO 16140. Additionally, a comparison between validated real-time PCR method and traditional methods based on the isolation of this pathogen on differential agar media was conducted. The validated method has shown the possibility of detection of 58 copies of ntnh gene (genome equivalents) for DNA obtained from dilution of pure Clostridium botulinum NCTC 887 cells. For DNA obtained from the contaminated food and feed samples inhibitory effect was observed. The tested method has shown high specificity proved by the examination of DNA obtained from C. botulinum reference strains and other strains of Clostridium sp. The specificity has also proved the obtained concordance between results from analyses using test on laboratory mice with those from analyses using the tested real-time PCR method. The obtained results have shown that the described method gives the possibility to detect the pathogen without isolation and to shorten time of analysis in comparison to the traditional methods, based on isolation of this pathogen on differential agar media.

19

Advanced methods of bacteriological identification in a clinical microbiology laboratory

75%

Zukowska M. E.

Journal of Pre-Clinical and Clinical Research

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2021

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tom 15

|

nr 2

20

Evaluation of fluorescence-based methods for total vs. amplifiable DNA quantification in plasma of lung cancer patients

75%

Szpechcinski A., Struniawski R., Zaleska J., Chabowski M., Orlowski T., Roszkowski K., Chorostowska-Wynimko J.

Journal of Physiology and Pharmacology. Supplement

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2008

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tom 59

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nr 6

675-681

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