scholarly journals Haemotropic Mycoplasma Infection Revealed by Real-Time PCR in Specific Pathogen-Free Rats

2014 ◽  
Vol 58 (3) ◽  
pp. 375-378 ◽  
Author(s):  
Hinako Sashida ◽  
Ryô Harasawa ◽  
Toshihiro Ichijo ◽  
Hiroshi Satoh ◽  
Kazuhisa Furuhama
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Real Time ◽  
Real Time Pcr ◽  
Wistar Rat ◽  
Rrna Gene ◽  
Specific Pathogen Free ◽  
Sprague Dawley ◽  
Laboratory Rats ◽  
Specific Pathogen

Abstract The presence of Mycoplasma haemomuris (haemoplasma) in blood samples collected from specific pathogen-free (SPF) laboratory rats bred in Japan was reported. Its presence was examined in Fischer 344, Sprague-Dawley (SD), and Wistar rat strains of both sexes by real-time PCR. All strains were positive for M. haemomuris infection. The 16S rRNA gene of M. haemomuris strain detected in the animals was amplified using end-point PCR. Only the entire nucleotide sequence of 16S rRNA gene of a mycoplasma strain detected in SD rats was determined and compared to those of other haemoplasmas. Our investigations suggest a wide M. haemomuris infection among the SPF rats purchased from commercial breeders in Japan.

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

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.

scholarly journals Use of 16S rRNA Gene-Targeted Group-Specific Primers for Real-Time PCR Analysis of Predominant Bacteria in Mouse Feces

2015 ◽  
Vol 81 (19) ◽  
pp. 6749-6756 ◽  
Author(s):  
Yun-Wen Yang ◽  
Mang-Kun Chen ◽  
Bing-Ya Yang ◽  
Xian-Jie Huang ◽  
Xue-Rui Zhang ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Real Time ◽  
Real Time Pcr ◽  
Bacterial Species ◽  
Chronic Inflammatory Bowel Disease ◽  
Trinitrobenzene Sulfonic Acid ◽  
Rrna Gene ◽  
Specific Primers ◽  
Content Type

ABSTRACTMouse models are widely used for studying gastrointestinal (GI) tract-related diseases. It is necessary and important to develop a new set of primers to monitor the mouse gut microbiota. In this study, 16S rRNA gene-targeted group-specific primers forFirmicutes,Actinobacteria,Bacteroidetes,Deferribacteres, “CandidatusSaccharibacteria,”Verrucomicrobia,Tenericutes, andProteobacteriawere designed and validated for quantification of the predominant bacterial species in mouse feces by real-time PCR. After confirmation of their accuracy and specificity by high-throughput sequencing technologies, these primers were applied to quantify the changes in the fecal samples from a trinitrobenzene sulfonic acid-induced colitis mouse model. Our results showed that this approach efficiently predicted the occurrence of colitis, such as spontaneous chronic inflammatory bowel disease in transgenic mice. The set of primers developed in this study provides a simple and affordable method to monitor changes in the intestinal microbiota at the phylum level.

scholarly journals Accurate and fast identification of Campylobacter fetus in bulls by real-time PCR targeting a 16S rRNA gene sequence

2020 ◽  
pp. 100163
Author(s):  
Rafael Delpiazzo ◽  
Maila Barcellos ◽  
Sofía Barros ◽  
Laura Betancor ◽  
Martín Fraga ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Real Time ◽  
Real Time Pcr ◽  
Gene Sequence ◽  
16S Rrna Gene Sequence ◽  
Rrna Gene ◽  
Rrna Gene Sequence ◽  
Fast Identification ◽  
Pcr Targeting

scholarly journals Simultaneous detection of 37 Lactobacillus species using a real-time PCR assay based on whole-genome sequence analysis

2020 ◽  
Author(s):  
Eiseul Kim ◽  
Seung-Min Yang ◽  
Bora Lim ◽  
Si Hong Park ◽  
Bryna Rackerby ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Real Time ◽  
Real Time Pcr ◽  
Genomic Analysis ◽  
23S Rrna ◽  
Comparative Genomic ◽  
Rrna Gene ◽  
Gene Sequences ◽  
Species Specific

Abstract BackgroundLactobacillus species are used as probiotics and play an important role in fermented food production. However, use of 16S rRNA gene sequences as standard markers for the differentiation of Lactobacillus species offers a very limited scope, as several species of Lactobacillus share similar 16S rRNA gene sequences. In this study, we developed a rapid and accurate method based on comparative genomic analysis for the simultaneous identification of 37 Lactobacillus species that are commonly used in probiotics and fermented foods. ResultsTo select species-specific sequences or genes, a total of 143 Lactobacillus complete genome sequences were compared using Python scripts. In 14 out of 37 species, species-specific sequences could not be found due to the similarity of the 16S–23S rRNA gene. Selected unique genes were obtained using comparative genomic analysis and all genes were confirmed to be specific for 52,478,804 genomes via in silico analysis; they were found not to be strain-specific, but to exist in all strains of the same species. Species-specific primer pairs were designed from the selected 16S–23S rRNA gene sequences or unique genes of species. The specificity of the species-specific primer pairs was confirmed using reference strains, and the accuracy and efficiency of the real-time polymerase chain reaction (PCR) with the standard curve were confirmed. The real-time PCR method developed in this study is able to accurately differentiate species that were not distinguishable using the 16S rRNA gene alone. This Real-time PCR method was designed to detect 37 Lactobacillus species in a single reaction. The developed method was then applied in the monitoring of 19 probiotics and 12 dairy products. The applied tests confirmed that the species detected in 17 products matched those indicated on their labels, whereas the remaining products contained species other than those appearing on the label. ConclusionsThe method developed in this study is able to rapidly and accurately distinguish different species of Lactobacillus, and can be used to monitor specific Lactobacillus species in foods such as probiotics and dairy products.

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