scholarly journals Invastigation of 16S rRNA, mecA and nuc genes in coagulase-positive and negative Staphylococci by Real-Time PCR

2021 ◽  
Author(s):  
Abdulbaki AKSAKAL ◽  
Şükrü ÖNALAN ◽  
Şeyda Şilan OKALİN
Keyword(s):  
16S Rrna ◽  
Real Time ◽  
Real Time Pcr

Determining Viability of M. ulcerans by 16S rRNA RT Reverse Transcriptase Real-Time PCR

2021 ◽  
pp. 81-86
Author(s):  
Marcus Beissner ◽  
Richard Odame Phillips ◽  
Gisela Bretzel
Keyword(s):  
16S Rrna ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Real Time Pcr

Monitoring of Microbial Community Inhabiting a Low-Grade Copper Sulphide Ore by Quantitative Real-Time PCR Analysis of 16S rRNA Genes

2007 ◽  
Vol 20-21 ◽  
pp. 539-542 ◽  
Author(s):  
Francisco Remonsellez ◽  
F. Galleguillos ◽  
Sonestie Janse van Rensburg ◽  
G.F. Rautenbach ◽  
Pedro A. Galleguillos ◽  
...  
Keyword(s):  
16S Rrna ◽  
Real Time ◽  
Real Time Pcr ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Low Grade ◽  
Copper Sulphide ◽  
Quantitative Real Time Pcr ◽  
Sulphide Ore ◽  
Heap Bioleaching

Microbial heap bioleaching is being used as an industrial process to recover copper from low grade ores. It is known that a consortium of different microorganisms participates in this process. Therefore identification and quantification of communities inhabiting heap bioleaching operations is a key step for understanding the dynamics and role of these microorganisms in the process. A quantitative real-time PCR approach was used to investigate the microbial dynamics in this process. To study the microbial population inhabiting a low-grade copper sulphide ore bioleaching industrial heap process at Escondida Mine in Chile, 16S rRNA genetic libraries were constructed using bacterial and archaeal universal primers. Phylogenetic analyses of sequences retrieved from genetic libraries showed that the community is mainly composed by microoganisms related to Acidithiobacillus ferrooxidans (2 strains), Acidithiobacillus thiooxidans, Leptospirillum ferrooxidans, Leptospirillum ferriphilum and the archaea Ferroplasma. Specific primers for real-time PCR determination were designed and tested to amplify each of the sequences obtained by cloning. Standard curves for real time PCR were performed using plasmid DNA from selected clones. This methodology is actually being used to monitor relevant microorganisms inhabiting this low-grade copper sulphide ore bioleaching industrial heap.

Real-time PCR of the 16S-rRNA gene in the diagnosis of neonatal bacteraemia

2008 ◽  
Vol 97 (10) ◽  
pp. 1376-1380 ◽  
Author(s):  
Andreas Ohlin ◽  
Anders Bäckman ◽  
Maria Björkqvist ◽  
Paula Mölling ◽  
Margaretha Jurstrand ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Real Time ◽  
Real Time Pcr ◽  
Rrna Gene ◽  
The 16S Rrna Gene

scholarly journals Prevalence of Swine Hemoplasmas Revealed by Real-Time PCR Using 16S rRNA Gene Primers

2012 ◽  
Vol 74 (10) ◽  
pp. 1315-1318 ◽  
Author(s):  
Yusaku WATANABE ◽  
Masatoshi FUJIHARA ◽  
Jin SUZUKI ◽  
Fumina SASAOKA ◽  
Kazuya NAGAI ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Real Time ◽  
Real Time Pcr ◽  
Rrna Gene

scholarly journals Usefulness of Multiplex Real-Time PCR for Simultaneous Pathogen Detection and Resistance Profiling of Staphylococcal Bacteremia

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Yousun Chung ◽  
Taek Soo Kim ◽  
Young Gi Min ◽  
Yun Ji Hong ◽  
Jeong Su Park ◽  
...  
Keyword(s):  
16S Rrna ◽  
Real Time ◽  
Real Time Pcr ◽  
Pathogen Detection ◽  
Methicillin Resistance ◽  
Blood Cultures ◽  
Pcr Assay ◽  
Accurate Identification ◽  
Essential Information ◽  
Methicillin Resistant

Staphylococci are the leading cause of nosocomial blood stream infections. Fast and accurate identification of staphylococci and confirmation of their methicillin resistance are crucial for immediate treatment with effective antibiotics. A multiplex real-time PCR assay that targetsmecA,femAspecific forS. aureus,femAspecific forS. epidermidis,16S rRNAfor universal bacteria, and16S rRNAspecific for staphylococci was developed and evaluated with 290 clinical blood culture samples containing Gram-positive cocci in clusters (GPCC). For the 262 blood cultures identified to the species level with the MicroScan WalkAway system (Siemens Healthcare Diagnostics, USA), the direct real-time PCR assay of positive blood cultures showed very good agreement for the categorization of staphylococci into methicillin-resistantS. aureus(MRSA), methicillin-susceptibleS. aureus(MSSA), methicillin-resistantS. epidermidis(MRSE), methicillin-susceptibleS. epidermidis(MSSE), methicillin-resistant non-S. epidermidisCoNS (MRCoNS), and methicillin-susceptible non-S. epidermidisCoNS (MSCoNS) (κ=0.9313). The direct multiplex real-time PCR assay of positive blood cultures containing GPCC can provide essential information at the critical point of infection with a turnaround time of no more than 4 h. Further studies should evaluate the clinical outcome of using this rapid real-time PCR assay in glycopeptide antibiotic therapy in clinical settings.

Impact of a probiotic, inulin, or their combination on the piglets’ microbiota at different intestinal locations

2015 ◽  
Vol 6 (4) ◽  
pp. 473-483 ◽  
Author(s):  
V.A. Sattler ◽  
K. Bayer ◽  
G. Schatzmayr ◽  
A.G. Haslberger ◽  
V. Klose
Keyword(s):  
16S Rrna ◽  
Gut Microbiota ◽  
16S Rrna Gene ◽  
Real Time ◽  
Relative Abundance ◽  
Real Time Pcr ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Feed Additives ◽  
Rrna Gene ◽  
16S Rrna Gene Pyrosequencing

Natural feed additives are used to maintain health and to promote performance of pigs without antibiotics. Effects of a probiotic, inulin, and their combination (synbiotic), on the microbial diversity and composition at different intestinal locations were analysed using denaturing gradient gel electrophoresis (DGGE), real-time PCR, and 16S rRNA gene pyrosequencing. Bacterial diversity assessed by DGGE and/or pyrosequencing was increased by inulin in all three gut locations and by the synbiotic in the caecum and colon. In contrast, the probiotic did only affect the microbiota diversity in the ileum. Shifts in the DGGE microbiota profiles of the caecum and colon were detected for the pro- and synbiotic fed animals, whereas inulin profiles were more similar to the ones of the control. 16S rRNA gene pyrosequencing revealed that all three additives could reduce Escherichia species in each gut location, indicating a potential beneficial effect on the gut microbiota. An increase of relative abundance of Clostridiaceae in the large intestine was found in the inulin group and of Enterococcaceae in the ileum of probiotic fed pigs. Furthermore, real-time PCR results showed that the probiotic and synbiotic increased bifidobacterial numbers in the ileum, which was supported by sequencing results. The probiotic and inulin, to different extents, changed the diversity, relative abundance of phylotypes, and community profiles of the porcine microbiota. However, alterations of the bacterial community were not uniformly between gut locations, demonstrating that functionality of feed additives is site specific. Therefore, gut sampling from various locations is crucial when investigations aim to identify the composition of a healthy gut microbiota after its manipulation through feed additives.

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