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original-paper | 25-August-2020

Establishment and Application of a Dual TaqMan Real-Time PCR Method for Proteus Mirabilis and Proteus Vulgaris

the technique has already been applied to the detection of P. mirabilis (Liu et al. 2019). At present, 16S ribosomal RNA (rRNA) sequencing, selective media, biochemical identification, and serological tests remain the mainstay modalities for distinguishing between strains of P. mirabilis and P. vulgaris (O’Hara et al. 2000). In this study, we used a dual TaqMan Real-Time PCR for the rapid and accurate identification and classification of P. mirabilis and P. vulgaris in food samples. Experimental

RUI YANG, GUOYANG XU, XIAOYOU WANG, ZHICHU QING, LIZHI FU

Polish Journal of Microbiology, Volume 69 , ISSUE 3, 293–300

original-paper | 08-September-2020

The Effect of Environmental Stresses on lipL32 Gene Expression in Pathogenic Leptospira spp. through Real-Time PCR

three isolates of Leptospira and the reference strain was isolated and purified with and without shock using a hybrid-RTM (Gene ALL Korea-South Seoul) RNA extraction kit. cDNA synthesis was performed by the TwoStep kit HyperScriptTM first-strand synthesis kit (Gene ALL Korea, South Seoul). cDNA was used as a template for PCR and real-time PCR. Real-Time PCR. The real-time PCR reaction was performed on Step One and Step One Plus Real-Time PCR systems (Applied Biosystems-Thermo Fisher Scientific

SONA ROSTAMPOUR YASOURI, MONIR DOUDI, MASOOD GHANE, NAFISEH SADAT NAGHAVI, ABOLHASAN REZAEI

Polish Journal of Microbiology, Volume 69 , ISSUE 3, 301–310

research-article | 30-November-2018

Developing a real-time PCR diagnostic method for a potential threat to chrysanthemum, Paratylenchus dianthus

. kumamotoensis are known to inhabit in many chrysanthemum growing areas (Iwahori et al., 2008) and Pratylenchus may cause chronic damage to chrysanthemum (Kobayashi, 1995). Chemical treatments including nematicides, such as fosthiazate, are popular means to control nematode damage in Okinawa, however, nematode diagnosis in a chrysanthemum field is not quick and easy. Koyama et al. (2016) reported a low cost and high-throughput approach to quantify Pratylenchus spp. with the real-time PCR method by using pre

Masanori Kawanobe, Koki Toyota, Hidehito Uchihara, Mikoto Takae

Journal of Nematology, Volume 51 , 1–11

Report | 12-March-2020

Application of real-time PCR and melting curve analysis in rapid Diego blood group genotyping

The paucity of appropriate reagents for serologic typing of the Diego blood group antigens has prompted the development of a real-time PCR and melting curve analysis for Diego blood group genotyping. In this study, we phenotyped 4326 donor blood samples for Dia using semiautomated equipment. All 157 Di(a+) samples were then genotyped by PCR using sequence-specific primers (PCR-SSP) for DI*02 because of anti-Dib scarcity. Of the 4326 samples, we simultaneously tested 160 samples for Dia and Dib

Marcia C. Zago Novaretti, Azulamara da Silva Ruiz, Pedro Enrique Dorlhiac-Llacer, Dalton Alencar Fisher Chamone

Immunohematology, Volume 26 , ISSUE 2, 66–70

Short Communication | 10-December-2018

Detection of Coxiella burnetii and Francisella tularensis in Tissues of Wild-living Animals and in Ticks of North-west Poland

Abstract This work presents results of the research on the occurrence of Coxiella burnetii and Francisella tularensis in the tissues of wild-living animals and ticks collected from Drawsko County, West Pomeranian Voivodeship. The real-time PCR testing for the pathogens comprised 928 samples of animal internal organs and 1551 ticks. The presence of C. burnetii was detected in 3% of wild-living animals and in 0.45–3.45% (dependent on collection areas) of ticks. The genetic sequences of F

AGATA BIELAWSKA-DRÓZD, PIOTR CIEŚLIK, DOROTA ŻAKOWSKA, PATRYCJA GŁOWACKA, BOŻENA WLIZŁO-SKOWRONEK, PRZEMYSŁAW ZIĘBA, ARKADIUSZ ZDUN

Polish Journal of Microbiology, Volume 67 , ISSUE 4, 529–534

Original Paper | 09-March-2018

Comparison of Methods Used for the Diagnosis of Epstein-Barr Virus Infections in Children

The accurate diagnosis of Epstein-Barr virus (EBV) infections is important, as many other infectious agents or diseases can cause similar symptoms. In this study, sera of pediatric patients who were suspected to have an EBV infection, were sent to Eskisehir Osmangazi University Faculty of Medicine, Department of Clinical Microbiology, and investigated by IFA, ELISA, immunoblotting and Real-time PCR. The performances of these tests were compared with IFA. The rates of agreement between ELISA and

Nilgun Kasifoglu, Semra Oz, Ener Cagri Dinleyici, Tercan Us, Ozcan Bor, Gul Durmaz, Yurdanur Akgun

Polish Journal of Microbiology, Volume 67 , ISSUE 1, 81–88

research-article | 30-November-2019

Molecular characterization of the Pratylenchus vulnus populations on cereals in Turkey

Vovlas, 2007). Molecular techniques as RAPD-PCR and sequencing of D2 to D3 expansion segments of the 28S rRNA was used for the identification of P. vulnus on different plant species (Subbotin et al., 2008; Bakooie et al., 2012; Lopez-Nicora et al., 2012). Moreover, real-time PCR provides sensitive identification of the species with species-specific primers using 1/128 of the DNA of one nematode (Huang and Yan, 2017). Pratylenchus vulnus (Allen and Jensen, 1951) (walnut root lesion nematode) has been

Mehmet Sait Karaca, Elif Yavuzaslanoglu, Gul Imriz, Ozlem Ates Sonmezoglu

Journal of Nematology, Volume 52 , 1–4

original-paper | 03-September-2019

Periodontal Status and Subgingival Biofilms in Cystic Fibrosis Adults

TAMARA PAWLACZYK-KAMIEŃSKA, RENATA ŚNIATAŁA, HALINA BATURA-GABRYEL, MARIA BORYSEWICZ-LEWICKA, SZCZEPAN COFTA

Polish Journal of Microbiology, Volume 68 , ISSUE 3, 377–382

Article | 14-October-2020

Assessment of the relative number of copies of the gene encoding human neutrophil antigen-2a (HNA-2a), CD177, and a homologous pseudogene by quantitative real-time PCR

from leukocytes of 12 subjects. The number of copies of exon 2 of CD177, an exon that is unique to this gene, and the number of copies of exon 9, an exon that is found in both CD177 and the pseudogene, was assessed with quantitative real-time PCR. The ratio of the number of copies of sequences homologous to CD177 exon 9 to the number of copies of exon 2 was 1.5 or greater in 7 of the 12 subjects, suggesting that both CD177 and the homologous pseudogene were present. The ratio of exon 9 to exon 2 in

Kristin Dittmar, Jong-Baeck Lim, Lorraine Caruccio, Maria Bettinotti, David Stroncek

Immunohematology, Volume 19 , ISSUE 4, 122–126

original-paper | 11-March-2020

Resensitization of Fluconazole-Resistant Urinary Candida spp. Isolates by Amikacin through Downregulation of Efflux Pump Genes

intervals of 0, 30, 60, 90 and 120 min, using a spectrofluorometer (Shimadzu, Japan) with excitation at 485 nm and emission at 530 nm. All results were represented as an average of three biological samples. Molecular quantification of the Candida efflux pump genes MDR1, CDR1, and CDR2 using quantitative real-time PCR. Quantitative real-time PCR was applied for two isolates C6 and C21 whose efflux pump activity was more prominently affected by amikacin using the Applied Biosystems 7500 Real-Time PCR

EVA A. EDWARD, NELLY M. MOHAMED, AZZA S. ZAKARIA

Polish Journal of Microbiology, Volume 69 , ISSUE 1, 73–84

Original Paper | 04-December-2017

Identification of Lactobacillus delbrueckii and Streptococcus thermophilus Strains Present in Artisanal Raw Cow Milk Cheese Using Real-time PCR and Classic Plate Count Methods

way and that genomic DNA solutions were free of PCR inhibitors. These methods revealed the presence of L. delbrueckii and S. thermophilus. The real-time PCR enabled quantification with a detection of 101–103 CFU/g of product. qPCR-standard curves were linear over seven log units down to 101 copies per reaction; efficiencies ranged from 77.9% to 93.6%. Cheese samples were analysed with plate count method and qPCR in parallel. Compared with the classic plate count method, the newly developed

Milena A. Stachelska

Polish Journal of Microbiology, Volume 66 , ISSUE 4, 491–499

Article | 14-October-2020

Rapid genotyping of the major alleles at the Duffy (FY) blood group locus using real-time fluorescence polymerase chain reaction

Fernando M. Araújo, Christina Pereira, Ana Aleixo, Isabel Henriques, Fátima Monteiro, Elsa Meireles, Pedro Lacerda, Luis M. Cunha-Ribeiro

Immunohematology, Volume 17 , ISSUE 2, 42–44

Article | 14-October-2020

Blood group antigen profile predicted by molecular biology - use of real-time polymerase chain reaction to genotype important KEL, JK, RHD, and RHCE alleles

Fernando Manuel Ferreira Araújo, Christiana Pereira, Fátima Monteiro, Isabel Henriques, Elsa Meireles, Pedro Lacerda, Ana Aleixo, Regina Celeste, Luis M. Cunha-Ribeiro, Maria J. Rodrigues

Immunohematology, Volume 18 , ISSUE 3, 59–64

Short Communication | 30-June-2018

Identification of Pathogenicity of Yersinia enterocolitica in Pig Tonsils Using the Real-Time PCR

MILENA A. STACHELSKA

Polish Journal of Microbiology, Volume 67 , ISSUE 2, –

original-paper | 28-June-2019

In situ Impact of the Antagonistic Fungal Strain, Trichoderma gamsii T30 on the Plant Pathogenic Fungus, Rhizoctonia solani in Soil

MUHAMMAD ANEES, MUHAMMAD ABID, SOBIA CHOHAN, MUHAMMAD JAMIL, NADEEM AHMED, LIXIN ZHANG, EUI SHIK RHA

Polish Journal of Microbiology, Volume 68 , ISSUE 2, 211–216

Report | 01-December-2019

Validation of a blood group genotyping method based on high-resolution melting curve analysis

Tianxiang Gong, Ying Hong, Naihong Wang, Xuemei Fu, Changhua Zhou

Immunohematology, Volume 30 , ISSUE 4, 161–165

short-communication | 30-November-2018

Comparison of Performance Characteristics of DxN VERIS System versus Qiagen PCR for HBV Genotype D and HCV Genotype 1b Quantification

. HCV PB analysis indicated DxN VERIS combined = −0.3394 + 0.8602 Qiagen log IU/ml with the correlation of 0.90. HCV plots for PB and BAP are shown in Fig. 2. Fig. 2. HCV plots for Passing-Bablok (upper) and Bland Altman analysis (lower). Viral nucleic acid detection is the gold standard for the detection of viral genomes in clinical samples. COBAS Ampliprep, artus Qiagen and Abbott real-time PCR assays are currently the most frequently used platforms worldwide in this field. DxN VERIS systems

MURAT SAYAN, AYSE ARIKAN, TAMER SANLIDAG

Polish Journal of Microbiology, Volume 68 , ISSUE 1, 139–143

Research Article | 22-May-2019

GENETIC DIFFERENTIATION METHODS OF MICROORGANISMS IN THE SOIL – PLANT SYSTEM

Małgorzata Łyszcz, Anna Gałązka

Postępy Mikrobiologii - Advancements of Microbiology, Volume 56 , ISSUE 3, 341–352

Report | 25-March-2020

Principles of PCR-based assays

DNA-based assays are powerful tools to predict the blood group of an individual and are rapidly gaining in popularity.  DNA, which can be extracted from various sources using commercial kits, is amplified by PCR to obtain a sufficient amount of the target of interest for analysis.  There are different types of PCR assays: standard single PCR (followed by RFLP or sequencing), allele-specific PCR, multiplex PCR, and real-time PCR.  Microarray platforms are a newer application of

Kim Hue-Roye, Sunitha Vege

Immunohematology, Volume 24 , ISSUE 4, 170–175

research-article | 06-April-2020

Investigating the synergic effects of valproic acid and crocin on BDNF and GDNF expression in epidermal neural crest stem cells

with the appropriate drug-containing medium. On the seventh day, total RNA was extracted from both control and treatment groups, cDNA was synthesized using the mRNA extracted from each sample, and real-time PCR was performed. Cell viability test To determine the non-toxic doses of VPA (Darou Pakhsh Pharma. Chem. Co, Iran) and crocin (Puyesh Darou Sina, Iran), an MTT assay was performed. EPI-NCSCs were seeded in 96-well plates, at a density of 5×103 cells/well, 24 h prior to treatment. Then, EPI

Zahra Baharvand, Mohammad Nabiuni, Mohammad Tahmaseb, Elaheh Amini, Sareh Pandamooz

Acta Neurobiologiae Experimentalis, Volume 80 , ISSUE 1, 38–46

original-paper | 18-February-2020

Impact of Hydrogen on the Transcriptome of Sinorhizobium meliloti 1021 Using RNA-sequencing Technology

RUIRUI LIU, LULU LI, ZHIYING LI, WEIWEI WANG

Polish Journal of Microbiology, Volume 69 , ISSUE 1, 39–48

Research Article | 20-May-2019

FRANCISELLA TULARENSIS – REVIEW 

. mediasiatica isolated mostly in Asia and F. tularensis subsp. novicida, non-pathogenic to humans. Due to its ability to infect and variable forms of the disease, the etiological agent of tularaemia is classified by the CDC (Centers for Disease Control and Prevention, USA) as a biological warfare agent with a high danger potential (group A). The majority of data describing incidence of tularaemia in Poland is based on serological tests. However, real-time PCR method and MST analysis of F. tularensis highly

Piotr Cieślik, Józef Knap, Agata Bielawska-Drózd

Postępy Mikrobiologii - Advancements of Microbiology, Volume 57 , ISSUE 1, 58–67

Original Paper | 10-December-2018

Comparison of PCR, Fluorescent in Situ Hybridization and Blood Cultures for Detection of Bacteremia in Children and Adolescents During Antibiotic Therapy

carried out in standard automated systems. Subsequently, FISH (Fluorescent In-Situ Hybridization) and nested multiplex-real-time-PCR (PCR) were performed. Blood cultures, FISH and PCR yielded positive results in 18%, 39.1%, and 71.7% of samples, respectively. Significant differences were found between the results obtained through culture before and after induction of antibiotherapy: 25.5% vs. 9.7%. There was no significant difference in FISH and PCR results in relation to antibiotics. The three

TOMASZ W. ŹRÓDŁOWSKI, DANUTA JURKIEWICZ-BADACZ, AGNIESZKA SROKA-OLEKSIAK, DOMINIKA SALAMON, MAŁGORZATA BULANDA, TOMASZ GOSIEWSKI

Polish Journal of Microbiology, Volume 67 , ISSUE 4, 479–486

research-article | 30-November-2020

Effect of fluensulfone on different functional genes of root-knot nematode Meloidogyne incognita

directly treating the nematode J2s with different concentrations of fluensulfone (10, 50, and 100 ppm), followed by analysis of transcript levels of the respective genes by quantitative real-time PCR (qRT PCR) at two time points (5 and 10 hr post exposure). Materials and methods Nematode population and drug material The pure culture of an Indian isolate of Meloidogyne incognita race 1 was raised on susceptible tomato plants (Solanum lycopersicum cv. Pusa ruby) in a glasshouse at ICAR-Indian

Alkesh Hada, Divya Singh, Kranti Kavalipurapu Veera Venkata Satyanarayana, Madhurima Chatterjee, Victor Phani, Uma Rao

Journal of Nematology, Volume 53 , 1–14

research-article | 30-June-2021

Effects of artemisinin and TSP-1-human endometrial-derived stem cells on a streptozocin-induced model of Alzheimer’s disease and diabetes in Wistar rats

isolated from leukocytes utilizing the RNX plusTM kit based on the manufacturer’s procedure (Cinnagen, Tehran, Iran). CDNA synthesis was performed by EasyTM cDNA Synthesis Kit (Parstous Biotechnology, Tehran, Iran) following the manufacturer’s instruction. Real-time PCR was carried out by a Bio-Rad Real-Time PCR detection system by utilizing SYBR green PCR master mix (Takara, Japan). Real-time PCR was adjusted in different three stages: first, initialization under the temperature of 95°C for 2 min

Poorgholam Parvin, Yaghmaei Parichehreh, Noureddini Mehdi, Hajebrahimi Zahra

Acta Neurobiologiae Experimentalis, Volume 81 , ISSUE 2, 141–150

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