The Experts below are selected from a list of 177 Experts worldwide ranked by ideXlab platform
Michael Kerin - One of the best experts on this subject based on the ideXlab platform.
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Development of a Molecular Methodology to quantify Staphylococcus epidermidis in surgical washout samples from prosthetic joint replacement surgery
European Journal of Orthopaedic Surgery & Traumatology, 2007Co-Authors: Fergus J. Byrne, Sinéad M. Waters, Peadar S. Waters, William Curtin, Michael KerinAbstract:L’infection prothétique est une complication sérieuse des arthroplasties totales total et entraîne une grande morbidité chez les patients qui en sont atteints La cause la plus commune des infections liées aux prothèses sont dues à diverses espèces de staphylocoque , y compris le staphylocoque epidermidis . La culture a servi de gold standard au diagnostic, en dépit des imperfections évidentes en termes de sensibilité et de temps. Des méthodologies géniquo-bactériennesd’extraction d’ADN ont été évaluées pour le rétablissement optimal de l’ADN génique des échantillons stérilisés de lavage, inoculés avec le staphylocoque epidermidis . Des analyses en temps réel de PCR visant le gène spécifique du gseA du staphylocoque epidermidis ont été conçues pour détecter de façon crertaine et pour quantifier le staphylocoque epidermidis . Soixante échantillons de produit de lavage pour des arthroplasties primaires de hanche et de genou ont été prélevés aseptiquement. Toutes les cultures ont étté trouvées stériles en utilisant l’approche culture-dépendante. Des échantillons ont été soumis à une analyse temps réel de PCR au staphylocoque epidermidis . La courbe standard a montré de bonnes linéarités. La limite de sensibilité de l’analyse était inférieure à CFU staphylocoque epidermidis:prélèvement . La reproductibilité de l’analyse a été confirmée. Le staphylocoque epidermidis n’a pas été retrouvé dans l’un quelconque d’entre ces échantillons en utilisant la technique nouvelle spécifique SYBR en temps réel de PCR. Les résultats ont indiqué que les produits de lavage étaient réellement négatifs et e contenaient pas du staphylocoque epidermidis .. Les patients n’ont d’ailleurs montré aucun symptôme d’infection, ce qui conforte cette affirmation. Pour illustrer la pleine efficacité de l’analyse en temps réel du roman PCR, un plus grand nombre d’échantillons doit être examiné (patients >1,000). Prosthetic joint infection is a serious complication of total joint arthroplasty that causes great morbidity in affected individuals. The most common cause of prosthesis-associated infections is members of Staphylococcus spp., including Staphylococcus epidermidis . Culture has served as the gold standard for diagnosis, despite obvious shortcomings in terms of sensitivity and time. Bacterial genomic DNA extraction methodologies were evaluated for optimal recovery of genomic DNA from sterilised washout samples, spiked with Staphylococcus epidermidis . Real time PCR assays targeting the Staphylococcus epidermidis specific gseA gene were designed to reliably detect and quantify S. epidermidis . Sixty postoperative washout samples from primary hip and knee arthroplasties were taken aseptically. All were shown to be culture negative using the culture-dependent approach. These samples were subjected to S. epidermidis -specific real time PCR. Standard curve showed good linearity. Sensitivity limit of the assay was less than 10 CFU S. epidermidis /sample. Reproducibility of the assay was confirmed. Staphylococcus epidermidis was not identified in any of the samples taken using the novel species-specific SYBR green real time PCR technique. Results indicated that washout samples were true negatives and did not harbour S. epidermidis . To support this, patients displayed no symptoms of infection. To illustrate the full effectiveness of the novel real time PCR assay, a larger number of samples need to be tested (>1,000 patients).
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Development of a Molecular Methodology to quantify Staphylococcus epidermidis in surgical wash-out samples from prosthetic joint replacement surgery
European journal of orthopaedic surgery & traumatology : orthopedie traumatologie, 2007Co-Authors: Fergus Byrne, Sinéad M. Waters, Peadar S. Waters, William Curtin, Michael KerinAbstract:Prosthetic joint infection is a serious complication of total joint arthroplasty that causes great morbidity in affected individuals. The most common cause of prosthesis associated infections are members of Staphylococcus spp., including Staphylococcus epidermidis. Culture has served as the gold standard for diagnosis, despite obvious shortcomings in terms of sensitivity and time. Bacterial genomic DNA extraction methodologies were evaluated for optimal recovery of genomic DNA from sterilised wash-out samples, spiked with S. epidermidis. Real time Polymerase chain reaction (PCR) assays targeting the S. epidermidis specific gseA gene were designed to reliably detect and quantify S. epidermidis. Sixty post-operative wash-out samples from primary hip and knee arthroplasties were taken aseptically. All were shown to be culture negative using the culture-dependent approach. These were samples were subjected to S. epidermidis-specific real time PCR. Standard curve showed good linearity. Sensitivity limit of the assay was 1,000 patients).
Richard C Hamelin - One of the best experts on this subject based on the ideXlab platform.
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Molecular assays to detect the presence and viability of phytophthora ramorum and grosmannia clavigera
PLOS ONE, 2020Co-Authors: Nicolas Feau, Barbara Wong, Isabel Leal, Angela Dale, Richard C Hamelin, Adnan UzunovicAbstract:Wood and wood products can harbor microorganisms that can raise phytosanitary concerns in countries importing or exporting these products. To evaluate the efficacy of wood treatment on the survival of microorganisms of phytosanitary concern the method of choice is to grow microbes in petri dishes for subsequent identification. However, some plant pathogens are difficult or impossible to grow in axenic cultures. A Molecular Methodology capable of detecting living fungi and fungus-like organisms in situ can provide a solution. RNA represents the transcription of genes and can become rapidly unstable after cell death, providing a proxy measure of viability. We designed and used RNA-based Molecular diagnostic assays targeting genes essential to vital processes and assessed their presence in wood colonized by fungi and oomycetes through reverse transcription and real-time polymerase chain reaction (PCR). A stability analysis was conducted by comparing the ratio of mRNA to gDNA over time following heat treatment of mycelial cultures of the Oomycete Phytophthora ramorum and the fungus Grosmannia clavigera. The real-time PCR results indicated that the DNA remained stable over a period of 10 days post treatment in heat-treated samples, whereas mRNA could not be detected after 24 hours for P. ramorum or 96 hours for G. clavigera. Therefore, this method provides a reliable way to evaluate the viability of these pathogens and offers a potential way to assess the effectiveness of existing and emerging wood treatments. This can have important phytosanitary impacts on assessing both timber and non-timber forest products of commercial value in international wood trade.
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Molecular assays to detect the presence and viability of phytophthora ramorum and grosmannia clavigera
bioRxiv, 2019Co-Authors: Nicolas Feau, Barbara Wong, Isabel Leal, Angela Dale, Richard C Hamelin, Adnan UzunovicAbstract:In order to determine if living microorganisms of phytosanitary concern are present in wood after eradication treatment and evaluate the efficacy of such treatments, the method of choice is to grow microbes in petri dishes for subsequent identification. However, some plant pathogens are difficult or impossible to grow in axenic cultures. A Molecular Methodology capable of detecting living fungi and fungus-like organisms in situ is required. Since RNA represents the transcription of genes, can therefore only be produced by living organisms and is rapidly degraded after cell death, it provides a proxy to determine if an organism is alive. RNA-based Molecular diagnostic assays targeting genes essential to vital processes in living fungi and fungi-like organisms were designed and used to assess their presence and viability in wood through reverse transcription and real-time polymerase chain reaction (PCR). A stability analysis was conducted by comparing the ratio of mRNA to gDNA over time following heat treatment of wood infected with the Oomycete Phytophthora ramorum and the fungus Grosmannia clavigera. The real-time PCR results indicated that for heat-treated samples, the DNA remained stable over a period of 10 days post treatment, whereas mRNA could not be detected after 24 hours for P. ramorum or 96 hours for G. clavigera. Therefore, this method provides a reliable way to evaluate viability of these pathogens and test the effectiveness of existing and emerging wood treatments. This can have important phytosanitary impacts on assessing both timber and non-timber forest products of commercial value in international wood trade.
Barbara Wong - One of the best experts on this subject based on the ideXlab platform.
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Molecular assays to detect the presence and viability of phytophthora ramorum and grosmannia clavigera
PLOS ONE, 2020Co-Authors: Nicolas Feau, Barbara Wong, Isabel Leal, Angela Dale, Richard C Hamelin, Adnan UzunovicAbstract:Wood and wood products can harbor microorganisms that can raise phytosanitary concerns in countries importing or exporting these products. To evaluate the efficacy of wood treatment on the survival of microorganisms of phytosanitary concern the method of choice is to grow microbes in petri dishes for subsequent identification. However, some plant pathogens are difficult or impossible to grow in axenic cultures. A Molecular Methodology capable of detecting living fungi and fungus-like organisms in situ can provide a solution. RNA represents the transcription of genes and can become rapidly unstable after cell death, providing a proxy measure of viability. We designed and used RNA-based Molecular diagnostic assays targeting genes essential to vital processes and assessed their presence in wood colonized by fungi and oomycetes through reverse transcription and real-time polymerase chain reaction (PCR). A stability analysis was conducted by comparing the ratio of mRNA to gDNA over time following heat treatment of mycelial cultures of the Oomycete Phytophthora ramorum and the fungus Grosmannia clavigera. The real-time PCR results indicated that the DNA remained stable over a period of 10 days post treatment in heat-treated samples, whereas mRNA could not be detected after 24 hours for P. ramorum or 96 hours for G. clavigera. Therefore, this method provides a reliable way to evaluate the viability of these pathogens and offers a potential way to assess the effectiveness of existing and emerging wood treatments. This can have important phytosanitary impacts on assessing both timber and non-timber forest products of commercial value in international wood trade.
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Molecular assays to detect the presence and viability of phytophthora ramorum and grosmannia clavigera
bioRxiv, 2019Co-Authors: Nicolas Feau, Barbara Wong, Isabel Leal, Angela Dale, Richard C Hamelin, Adnan UzunovicAbstract:In order to determine if living microorganisms of phytosanitary concern are present in wood after eradication treatment and evaluate the efficacy of such treatments, the method of choice is to grow microbes in petri dishes for subsequent identification. However, some plant pathogens are difficult or impossible to grow in axenic cultures. A Molecular Methodology capable of detecting living fungi and fungus-like organisms in situ is required. Since RNA represents the transcription of genes, can therefore only be produced by living organisms and is rapidly degraded after cell death, it provides a proxy to determine if an organism is alive. RNA-based Molecular diagnostic assays targeting genes essential to vital processes in living fungi and fungi-like organisms were designed and used to assess their presence and viability in wood through reverse transcription and real-time polymerase chain reaction (PCR). A stability analysis was conducted by comparing the ratio of mRNA to gDNA over time following heat treatment of wood infected with the Oomycete Phytophthora ramorum and the fungus Grosmannia clavigera. The real-time PCR results indicated that for heat-treated samples, the DNA remained stable over a period of 10 days post treatment, whereas mRNA could not be detected after 24 hours for P. ramorum or 96 hours for G. clavigera. Therefore, this method provides a reliable way to evaluate viability of these pathogens and test the effectiveness of existing and emerging wood treatments. This can have important phytosanitary impacts on assessing both timber and non-timber forest products of commercial value in international wood trade.
Fergus J. Byrne - One of the best experts on this subject based on the ideXlab platform.
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Development of a Molecular Methodology to quantify Staphylococcus epidermidis in surgical washout samples from prosthetic joint replacement surgery
European Journal of Orthopaedic Surgery & Traumatology, 2007Co-Authors: Fergus J. Byrne, Sinéad M. Waters, Peadar S. Waters, William Curtin, Michael KerinAbstract:L’infection prothétique est une complication sérieuse des arthroplasties totales total et entraîne une grande morbidité chez les patients qui en sont atteints La cause la plus commune des infections liées aux prothèses sont dues à diverses espèces de staphylocoque , y compris le staphylocoque epidermidis . La culture a servi de gold standard au diagnostic, en dépit des imperfections évidentes en termes de sensibilité et de temps. Des méthodologies géniquo-bactériennesd’extraction d’ADN ont été évaluées pour le rétablissement optimal de l’ADN génique des échantillons stérilisés de lavage, inoculés avec le staphylocoque epidermidis . Des analyses en temps réel de PCR visant le gène spécifique du gseA du staphylocoque epidermidis ont été conçues pour détecter de façon crertaine et pour quantifier le staphylocoque epidermidis . Soixante échantillons de produit de lavage pour des arthroplasties primaires de hanche et de genou ont été prélevés aseptiquement. Toutes les cultures ont étté trouvées stériles en utilisant l’approche culture-dépendante. Des échantillons ont été soumis à une analyse temps réel de PCR au staphylocoque epidermidis . La courbe standard a montré de bonnes linéarités. La limite de sensibilité de l’analyse était inférieure à CFU staphylocoque epidermidis:prélèvement . La reproductibilité de l’analyse a été confirmée. Le staphylocoque epidermidis n’a pas été retrouvé dans l’un quelconque d’entre ces échantillons en utilisant la technique nouvelle spécifique SYBR en temps réel de PCR. Les résultats ont indiqué que les produits de lavage étaient réellement négatifs et e contenaient pas du staphylocoque epidermidis .. Les patients n’ont d’ailleurs montré aucun symptôme d’infection, ce qui conforte cette affirmation. Pour illustrer la pleine efficacité de l’analyse en temps réel du roman PCR, un plus grand nombre d’échantillons doit être examiné (patients >1,000). Prosthetic joint infection is a serious complication of total joint arthroplasty that causes great morbidity in affected individuals. The most common cause of prosthesis-associated infections is members of Staphylococcus spp., including Staphylococcus epidermidis . Culture has served as the gold standard for diagnosis, despite obvious shortcomings in terms of sensitivity and time. Bacterial genomic DNA extraction methodologies were evaluated for optimal recovery of genomic DNA from sterilised washout samples, spiked with Staphylococcus epidermidis . Real time PCR assays targeting the Staphylococcus epidermidis specific gseA gene were designed to reliably detect and quantify S. epidermidis . Sixty postoperative washout samples from primary hip and knee arthroplasties were taken aseptically. All were shown to be culture negative using the culture-dependent approach. These samples were subjected to S. epidermidis -specific real time PCR. Standard curve showed good linearity. Sensitivity limit of the assay was less than 10 CFU S. epidermidis /sample. Reproducibility of the assay was confirmed. Staphylococcus epidermidis was not identified in any of the samples taken using the novel species-specific SYBR green real time PCR technique. Results indicated that washout samples were true negatives and did not harbour S. epidermidis . To support this, patients displayed no symptoms of infection. To illustrate the full effectiveness of the novel real time PCR assay, a larger number of samples need to be tested (>1,000 patients).
Sinéad M. Waters - One of the best experts on this subject based on the ideXlab platform.
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Development of a Molecular Methodology to quantify Staphylococcus epidermidis in surgical washout samples from prosthetic joint replacement surgery
European Journal of Orthopaedic Surgery & Traumatology, 2007Co-Authors: Fergus J. Byrne, Sinéad M. Waters, Peadar S. Waters, William Curtin, Michael KerinAbstract:L’infection prothétique est une complication sérieuse des arthroplasties totales total et entraîne une grande morbidité chez les patients qui en sont atteints La cause la plus commune des infections liées aux prothèses sont dues à diverses espèces de staphylocoque , y compris le staphylocoque epidermidis . La culture a servi de gold standard au diagnostic, en dépit des imperfections évidentes en termes de sensibilité et de temps. Des méthodologies géniquo-bactériennesd’extraction d’ADN ont été évaluées pour le rétablissement optimal de l’ADN génique des échantillons stérilisés de lavage, inoculés avec le staphylocoque epidermidis . Des analyses en temps réel de PCR visant le gène spécifique du gseA du staphylocoque epidermidis ont été conçues pour détecter de façon crertaine et pour quantifier le staphylocoque epidermidis . Soixante échantillons de produit de lavage pour des arthroplasties primaires de hanche et de genou ont été prélevés aseptiquement. Toutes les cultures ont étté trouvées stériles en utilisant l’approche culture-dépendante. Des échantillons ont été soumis à une analyse temps réel de PCR au staphylocoque epidermidis . La courbe standard a montré de bonnes linéarités. La limite de sensibilité de l’analyse était inférieure à CFU staphylocoque epidermidis:prélèvement . La reproductibilité de l’analyse a été confirmée. Le staphylocoque epidermidis n’a pas été retrouvé dans l’un quelconque d’entre ces échantillons en utilisant la technique nouvelle spécifique SYBR en temps réel de PCR. Les résultats ont indiqué que les produits de lavage étaient réellement négatifs et e contenaient pas du staphylocoque epidermidis .. Les patients n’ont d’ailleurs montré aucun symptôme d’infection, ce qui conforte cette affirmation. Pour illustrer la pleine efficacité de l’analyse en temps réel du roman PCR, un plus grand nombre d’échantillons doit être examiné (patients >1,000). Prosthetic joint infection is a serious complication of total joint arthroplasty that causes great morbidity in affected individuals. The most common cause of prosthesis-associated infections is members of Staphylococcus spp., including Staphylococcus epidermidis . Culture has served as the gold standard for diagnosis, despite obvious shortcomings in terms of sensitivity and time. Bacterial genomic DNA extraction methodologies were evaluated for optimal recovery of genomic DNA from sterilised washout samples, spiked with Staphylococcus epidermidis . Real time PCR assays targeting the Staphylococcus epidermidis specific gseA gene were designed to reliably detect and quantify S. epidermidis . Sixty postoperative washout samples from primary hip and knee arthroplasties were taken aseptically. All were shown to be culture negative using the culture-dependent approach. These samples were subjected to S. epidermidis -specific real time PCR. Standard curve showed good linearity. Sensitivity limit of the assay was less than 10 CFU S. epidermidis /sample. Reproducibility of the assay was confirmed. Staphylococcus epidermidis was not identified in any of the samples taken using the novel species-specific SYBR green real time PCR technique. Results indicated that washout samples were true negatives and did not harbour S. epidermidis . To support this, patients displayed no symptoms of infection. To illustrate the full effectiveness of the novel real time PCR assay, a larger number of samples need to be tested (>1,000 patients).
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Development of a Molecular Methodology to quantify Staphylococcus epidermidis in surgical wash-out samples from prosthetic joint replacement surgery
European journal of orthopaedic surgery & traumatology : orthopedie traumatologie, 2007Co-Authors: Fergus Byrne, Sinéad M. Waters, Peadar S. Waters, William Curtin, Michael KerinAbstract:Prosthetic joint infection is a serious complication of total joint arthroplasty that causes great morbidity in affected individuals. The most common cause of prosthesis associated infections are members of Staphylococcus spp., including Staphylococcus epidermidis. Culture has served as the gold standard for diagnosis, despite obvious shortcomings in terms of sensitivity and time. Bacterial genomic DNA extraction methodologies were evaluated for optimal recovery of genomic DNA from sterilised wash-out samples, spiked with S. epidermidis. Real time Polymerase chain reaction (PCR) assays targeting the S. epidermidis specific gseA gene were designed to reliably detect and quantify S. epidermidis. Sixty post-operative wash-out samples from primary hip and knee arthroplasties were taken aseptically. All were shown to be culture negative using the culture-dependent approach. These were samples were subjected to S. epidermidis-specific real time PCR. Standard curve showed good linearity. Sensitivity limit of the assay was 1,000 patients).