Evaluation of nanopore sequencing as a diagnostic tool for Mycoplasma bovis
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PROVIDER: PRJEB47272 | ENA |
REPOSITORIES: ENA
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Project description:Rapid identification of Mycoplasma bovis infections in cattle is a key factor to guide antimicrobial therapy and biosecurity measures. Recently, Nanopore sequencing became an affordable diagnostic tool for both clinically relevant viruses and bacteria, but the diagnostic accuracy for M. bovis identification is undocumented. Therefore, in this study Nanopore sequencing was compared to rapid identification of M. bovis with matrix-assisted laser desorption ionization-time of flight mass spectrometry (RIMM) and a triplex real-time PCR assay in a Bayesian latent class model (BLCM) for M. bovis in bronchoalveolar lavage fluid (BALf) samples obtained from calves. In practice, pooling of samples is often used to save money, but the influence on diagnostic accuracy has not been described for M. bovis. Therefore, a convenience sample of 17 pooled samples containing 5 individual BALf samples per farm was analyzed as well. The results for the pooled samples were compared with those for the individual samples to determine sensitivity and specificity. The BLCM showed good sensitivity (77.3% [95% credible interval, 57.8 to 92.8%]) and high specificity (97.4% [91.5 to 99.7%]) for Nanopore sequencing, compared to RIMM (sensitivity, 93.0% [76.8 to 99.5%]; specificity, 91.3% [82.5 to 97.0%]) and real-time PCR (sensitivity, 94.6% [89.7 to 97.7%]; specificity, 86.0% [76.1 to 93.6%]). Sensitivity and specificity of pooled analysis for M. bovis were 85.7% (95% confidence interval, 59.8 to 111.6%) and 90.0% (71.4 to 108.6%%), respectively, for Nanopore sequencing and 100% (100% to 100%) and 88.9% (68.4 to 109.4%) for RIMM. In conclusion, Nanopore sequencing is a rapid, reliable tool for the identification of M. bovis. To reduce costs and increase the chance of M. bovis identification, pooling of 5 samples for Nanopore sequencing and RIMM is possible.
| S-EPMC8601226 | biostudies-literature
Project description:This study was aimed to elucidate a global antigenic profile of Mycoplasma bovis (M. bovis) with immunoproteomics, immunoinformatics, and gene expression approaches. The extracts of whole-cell proteins and TX-114 membrane fraction of a Chinese strain M. bovis HB0801 were separated with two dimensional gel electrophoresis (2-DE) and proteins reacting with antisera to M. bovis from experimentally infected calves were detected by MALDI-TOF MS.
2016-06-13 | PXD003479 | Pride
Project description:The immune response associated with mastitis caused by Mycoplasma bovis is a very complicated biological process in several type of cells, including immune cells, mammary epithelial cells and, endothelial cells. Thus, revealing of the microRNAs in the Mycoplasma bovis infected mammary gland tissues is particularly important for the immune response mechanism to Mycoplasma bovis. Firstly, mammary gland tissue samples were collected from Holstein cows and screened for Mycoplasma bovis. Then, total RNA was isolated from mycoplasma bovis infected tissues and RNA sequencing was performed. After bioinformatics analysis, GO and KEGG analysis of target genes of identified microRNAs were conducted. Our results revaled that 24 of the known microRNAs were expressed differently and 13 of the novel microRNAs were expressed differently in Mycoplasma bovis positive tissues. The target genes of these microRNAs were found to be associated with especially inflammation pathways. In conclusion, this study demonstrated that identified miRNAs may be involved in the signaling pathways during mastitis case caused by Mycoplasma bovis.
2021-05-25 | GSE152865 | GEO
Project description:Comparing differential expressed proteins between Mycoplasma bovis different strains by using label-free method
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Project description:Mycoplasma bovis (M. bovis) causes to the diseases such as arthritis, pneumonia, abortion and mastitis causing to great losses in dairy bovine industries. It undertakes significant functions in the regulation of the immune response formed by many RNA type bacteria such as Messenger RNA (mRNAs) and Long noncoding RNA (lncRNAs). mRNA and lncRNA expression profiles still cannot be majorly known in the bovine mammary gland tissues infected with M. bovis. To illuminate this issue, transcription analysis has been conducted regarding mRNA and IncRNAs in the breast tissues belonging to Holstein cattle infected and not infected with M. Bovis. Differently expressed 1310 mRNAs and 57 IncRNAs have been determined in the bovine mammary gland tissues infected and not infected with M. Bovis. In addition; 392 novel lncRNAs have been detected. Also; it has been found that 19 novel lncRNAs are expressed in a different way. It has been revealed in gene ontology analysis that the differently expressed mRNAs and IncRNAs play a significant role in many vital biological pathways such as Metabolic pathways, T cell receptor signaling pathway, TGF-beta signaling pathway, Pathways in cancer, PI3K-Akt signaling pathway, NF-kappa B signaling pathway, mTOR signaling pathway and Apoptosis including the immune response and cancer. When literature review is conducted, it is seen that this study is the first genome wide IncRNA research conducted on the infected bovine mammary gland tissues with M. bovis agent. Our results have provided bovine mammary gland IncRNA and mRNA resource to understand their roles in the regulation of the immune response against the agent of M. Bovis in the bovine mammary gland tissues.
2020-06-01 | GSE133333 | GEO
Project description:Mycoplasma species are highly contagious pathogens, and Mycoplasma infectious disease are a serious issue for the dairy industry. The bovine neutrophils play an important role for the eradication of pathogens which cause mycoplasmal infection, however the effects of M. bovis for immune response of neutrophils have not been fully clarified. We examined the transcription profiling of bovine neutrophils on the stimulation with M. bovis for 3h (3 stimuli, 3 control).
2020-11-30 | E-MTAB-9022 | biostudies-arrayexpress
Project description:Mycoplasma bovis is one of the major causative pathogens of the bovine respiratory complex disease that is characterized by enzootic pneumonia, mastitis, pleuritis and polyarthritis. M. bovis enters and colonizes the bovine respiratory epithelia through inhalation of aerosol from contaminated air. The nature of the interaction between M. bovis and bovine innate immune system is not well understood. We hypothesized that M. bovis invades blood monocytes and regulates cellular function to support its persistence and systemic dissemination. We used bovine-specific peptide kinome arrays to identify cellular signaling pathways that could be relevant to M. bovis-monocyte interaction in vitro. We validated these pathways using functional, protein and gene expression arrays. Here we show that infection of blood monocytes with M. bovis delays spontaneous or TNF-α/staurosporine-driven apoptosis, activates NF-κβ p65 subunit and inhibits caspase-9 activity. We also report that M. bovis infected bovine monocytes do not produce IFN-γ and TNF-α, although production of IL-10 is elevated. Our findings suggest that M. bovis takes over the cellular machinery of bovine monocytes to prolong bacterial survival and to possibly facilitate subsequent systemic distribution.
2016-06-07 | GSE47743 | GEO
Project description:Cyclic di-AMP is an essential and ubiquitous second messenger that regulates bacterial potassium (K+) concentrations to maintain osmotic equilibrium. And its specific K+ regulation mechanism in Mycoplasma has rarely been investigated. We used the ruminant pathogen Mycoplasma bovis (M. bovis) to investigate this mechanism. We verified that MbovP496 is a c-di-AMP synthetase and its mutant T5.415 showed growth inhibition under high K+ condition. To deliver why the mutant strain T5.415 can regulate M. bovis growth, we held a RNA-seq analysis to reveal that the differentially expressed genes between WT and T5.415 under normal condition and high K+ condition. The K+ transport pathways and metabolism related pathways were enriched. In conclusion, we found that the mutation of c-di-AMP synthetase, MbovP496 can broadly influence not only the metal ions transport pathway, but also has an effect on metabolism pathways,which could significantly contribute to understanding the regulation of growth by c-di-AMP synthetase in mycoplasmas.
2024-12-01 | GSE282920 | GEO
Project description:Mycoplasma species are highly contagious pathogens, and Mycoplasma infectious disease are a serious issue for the dairy industry. The bovine peripheral blood mononuclear cells play an important role for mycoplasma mastitis, however, the effects of M. bovis for immune response of peripheral blood mononuclear cells have not been fully clarified.We examined the transcription profiling of bovine peripheral blood mononuclear cells in intramammary infusion of M. bovis at day 7.
2019-12-19 | E-MTAB-7905 | biostudies-arrayexpress