The Experts below are selected from a list of 466068 Experts worldwide ranked by ideXlab platform
Bernard La Scola - One of the best experts on this subject based on the ideXlab platform.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled to high content Screening
Clinical Microbiology and Infection, 2021Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:Abstract Objectives A novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is responsible for the current coronavirus disease 2019 global pandemic. Only a few laboratories routinely isolate the virus, which is because the current co-culture strategy is highly time-consuming and requires a biosafety level 3 laboratory. This work aimed to develop a new high-throughput isolation strategy using novel technologies for rapid and automated isolation of SARS-CoV-2. Methods We used an automated microscope based on High-Content Screening (HCS), and we applied specific image analysis algorithms targeting cytopathic effects of SARS-CoV-2 on Vero E6 cells. A randomized panel of 104 samples, including 72 that tested positive by RT-PCR and 32 that tested negative, were processed with our HCS strategy and were compared with the classical isolation procedure. Results The isolation rate was 43% (31/72) with both strategies on RT-PCR-positive samples and was correlated with the initial RNA viral load in the samples, in which we obtained a positivity threshold of 27 Ct. Co-culture delays were shorter with the HCS strategy, where 80% (25/31) of the positive samples were recovered by the third day of co-culture, compared with only 26% (8/30) with the classic strategy. Moreover, only the HCS strategy allowed us to recover all the positive samples (31 with HCS versus 27 with classic strategy) after 1 week of co-culture. Conclusions This system allows the rapid and automated Screening of clinical samples with minimal operator workload, which reduces the risk of contamination and paves the way for future applications in clinical microbiology, such as large-scale drug susceptibility testing.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled with high content Screening
bioRxiv, 2020Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:SARS-CoV-2, a novel coronavirus infecting humans, is responsible for the current COVID-19 global pandemic. If several strains could be isolated worldwide, especially for in-vitro drug susceptibility testing and vaccine development, few laboratories routinely isolate SARS-CoV-2. This is due to the fact that the current co-culture strategy is highly time consuming and requires working in a biosafety level 3 laboratory. In this work, we present a new strategy based on high content Screening automated microscopy (HCS) allowing large scale isolation of SARS-CoV-2 from clinical samples in 1 week. A randomized panel of 104 samples, including 72 tested positive by RT-PCR and 32 tested negative, were processed with our HCS procedure and were compared to the classical isolation procedure. Isolation rate was 43% with both strategies on RT-PCR positive samples, and was correlated with the initial RNA viral load in the samples, where we obtained a positivity threshold of 27Ct. Co-culture delays were shorter with HCS strategy, where 80% of the positive samples were recovered by the third day of co-culture, as compared to only 25% with the classic strategy. Moreover, only the HCS strategy allowed us to recover all the positive elements after 1 week of co-culture. This system allows rapid and automated Screening of clinical samples with minimal operator intervention, thus accelerating the work rate and minimizing the risk of contamination of the laboratory personnel.
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high content Screening a reliable system for coxiella burnetii isolation from clinical samples
Journal of Clinical Microbiology, 2020Co-Authors: Rania Francis, Maxime Mioulane, Marion Le Bideau, Mariecharlotte Mati, Jacques Yaacoub Bou Khalil, Pierre-edouard Fournier, Bernard La ScolaAbstract:ABSTRACT Q fever, caused by Coxiella burnetii, is a worldwide zoonotic disease that may cause severe forms in humans and requires a specific and prolonged antibiotic treatment. Although current serological and molecular detection tools allow a reliable diagnosis of the disease, culture of C. burnetii strains is mandatory to assess their susceptibility to antibiotics and sequence their genome in order to optimize patient management and epidemiological studies. However, cultivating this fastidious microorganism is difficult and restricted to reference centers, as it requires biosafety level 3 laboratories and relies on cell culture performed by experienced technicians. In addition, the culture yield is low, which results in a small number of isolates being available. In this work, we developed a novel High-Content Screening (HCS) isolation strategy based on optimized high-throughput cell culture and automated microscopic detection of infected cells with specifically designed algorithms targeting cytopathic effects. This method was more efficient than the shell vial assay, at the level of time dependency, when applied to both frozen specimens (7 isolates recovered by HCS only, sensitivity 91% versus 78% for shell vial) and fresh samples (1 additional isolate using HCS, sensitivity 7% versus 5% for shell vial), for which most strains were recovered more rapidly with the new technique. In addition, detecting positive cultures by an automated microscope reduced the need for expertise and saved 24% of technician working time. Application of HCS to antibiotic susceptibility testing of 12 strains demonstrated that it was as efficient as the standard procedure that combines shell vial culture and quantitative PCR.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled to high content Screening
Clinical Microbiology and Infection, 2020Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:Objectives A novel coronavirus, SARS-CoV-2, is responsible for the current COVID-19 global pandemic Only a few laboratories routinely isolate the virus, which is because the current co-culture strategy is highly time-consuming and requires working in a biosafety level 3 laboratory This work aimed to develop a new high-throughput isolation strategy using novel technologies for rapid and automated isolation of SARS-CoV-2 Methods We used an automated microscope based on High-Content Screening (HCS), and we applied specific image analysis algorithms targeting cytopathic effects of SARS-CoV-2 on Vero E6 cells A randomized panel of 104 samples, including 72 that tested positive by RT-PCR and 32 that tested negative, were processed with our HCS strategy and were compared to the classical isolation procedure Results The isolation rate was 43% (31/72) with both strategies on RT-PCR-positive samples and was correlated with the initial RNA viral load in the samples, in which we obtained a positivity threshold of 27 Ct Co-culture delays were shorter with the HCS strategy, where 80% (25/31) of the positive samples were recovered by the third day of co-culture, compared to only 26% (8/30) with the classic strategy Moreover, only the HCS strategy allowed us to recover all the positive samples (31 with HCS versus 27 with classic strategy) after 1 week of co-culture Conclusions This system allows the rapid and automated Screening of clinical samples with minimal operator workload, which reduces the risk of contamination, thus paving the way for future applications in clinical microbiology, such as large-scale drug susceptibility testing
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high content Screening a reliable system for coxiella burnetii isolation from clinical samples
bioRxiv, 2019Co-Authors: Rania Francis, Maxime Mioulane, Marion Le Bideau, Mariecharlotte Mati, Jacques Bou Khalil, Pierre-edouard Fournier, Didier Raoult, Bernard La ScolaAbstract:Q fever, caused by Coxiella burnetii, is a worldwide zoonotic disease that may cause severe forms in humans and requires a specific and prolonged antibiotic treatment. Although the current serological and molecular detection tools enable a reliable diagnosis of the disease, culture of C. burnetii strains is mandatory to evaluate their antibiotic susceptibility and sequence their genome in order to optimize patient management and epidemiological studies. However, cultivating this fastidious microorganism is difficult and restricted to reference centers as it requires biosafety-level 3 laboratories and relies on cell culture performed by experienced technicians. In addition, the culture yield is low, which results in a small number of isolates being available. In this work, we developed a novel high content Screening (HCS) isolation strategy based on optimized high-throughput cell culture and automated microscopic detection of infected cells with specifically-designed algorithms targeting cytopathic effects. This method was more efficient than the shell-vial assay when applied to both frozen specimens (7 isolates recovered by HCS only, sensitivity 91% vs 78% for shell-vial) and fresh samples (1 additional isolate using HCS, sensitivity 7% vs 5% for shell-vial). In addition, detecting positive cultures by an automated microscope reduced the need for expertise and saved 24% of technician working time. Application of HCS to antibiotic susceptibility testing of 12 strains demonstrated that it was as efficient as the standard procedure that combines shell-vial culture and quantitative PCR. Overall, this high-throughput HCS system paves the way to the development of improved cell culture isolation of human viruses.
Rania Francis - One of the best experts on this subject based on the ideXlab platform.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled to high content Screening
Clinical Microbiology and Infection, 2021Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:Abstract Objectives A novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is responsible for the current coronavirus disease 2019 global pandemic. Only a few laboratories routinely isolate the virus, which is because the current co-culture strategy is highly time-consuming and requires a biosafety level 3 laboratory. This work aimed to develop a new high-throughput isolation strategy using novel technologies for rapid and automated isolation of SARS-CoV-2. Methods We used an automated microscope based on High-Content Screening (HCS), and we applied specific image analysis algorithms targeting cytopathic effects of SARS-CoV-2 on Vero E6 cells. A randomized panel of 104 samples, including 72 that tested positive by RT-PCR and 32 that tested negative, were processed with our HCS strategy and were compared with the classical isolation procedure. Results The isolation rate was 43% (31/72) with both strategies on RT-PCR-positive samples and was correlated with the initial RNA viral load in the samples, in which we obtained a positivity threshold of 27 Ct. Co-culture delays were shorter with the HCS strategy, where 80% (25/31) of the positive samples were recovered by the third day of co-culture, compared with only 26% (8/30) with the classic strategy. Moreover, only the HCS strategy allowed us to recover all the positive samples (31 with HCS versus 27 with classic strategy) after 1 week of co-culture. Conclusions This system allows the rapid and automated Screening of clinical samples with minimal operator workload, which reduces the risk of contamination and paves the way for future applications in clinical microbiology, such as large-scale drug susceptibility testing.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled with high content Screening
bioRxiv, 2020Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:SARS-CoV-2, a novel coronavirus infecting humans, is responsible for the current COVID-19 global pandemic. If several strains could be isolated worldwide, especially for in-vitro drug susceptibility testing and vaccine development, few laboratories routinely isolate SARS-CoV-2. This is due to the fact that the current co-culture strategy is highly time consuming and requires working in a biosafety level 3 laboratory. In this work, we present a new strategy based on high content Screening automated microscopy (HCS) allowing large scale isolation of SARS-CoV-2 from clinical samples in 1 week. A randomized panel of 104 samples, including 72 tested positive by RT-PCR and 32 tested negative, were processed with our HCS procedure and were compared to the classical isolation procedure. Isolation rate was 43% with both strategies on RT-PCR positive samples, and was correlated with the initial RNA viral load in the samples, where we obtained a positivity threshold of 27Ct. Co-culture delays were shorter with HCS strategy, where 80% of the positive samples were recovered by the third day of co-culture, as compared to only 25% with the classic strategy. Moreover, only the HCS strategy allowed us to recover all the positive elements after 1 week of co-culture. This system allows rapid and automated Screening of clinical samples with minimal operator intervention, thus accelerating the work rate and minimizing the risk of contamination of the laboratory personnel.
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high content Screening a reliable system for coxiella burnetii isolation from clinical samples
Journal of Clinical Microbiology, 2020Co-Authors: Rania Francis, Maxime Mioulane, Marion Le Bideau, Mariecharlotte Mati, Jacques Yaacoub Bou Khalil, Pierre-edouard Fournier, Bernard La ScolaAbstract:ABSTRACT Q fever, caused by Coxiella burnetii, is a worldwide zoonotic disease that may cause severe forms in humans and requires a specific and prolonged antibiotic treatment. Although current serological and molecular detection tools allow a reliable diagnosis of the disease, culture of C. burnetii strains is mandatory to assess their susceptibility to antibiotics and sequence their genome in order to optimize patient management and epidemiological studies. However, cultivating this fastidious microorganism is difficult and restricted to reference centers, as it requires biosafety level 3 laboratories and relies on cell culture performed by experienced technicians. In addition, the culture yield is low, which results in a small number of isolates being available. In this work, we developed a novel High-Content Screening (HCS) isolation strategy based on optimized high-throughput cell culture and automated microscopic detection of infected cells with specifically designed algorithms targeting cytopathic effects. This method was more efficient than the shell vial assay, at the level of time dependency, when applied to both frozen specimens (7 isolates recovered by HCS only, sensitivity 91% versus 78% for shell vial) and fresh samples (1 additional isolate using HCS, sensitivity 7% versus 5% for shell vial), for which most strains were recovered more rapidly with the new technique. In addition, detecting positive cultures by an automated microscope reduced the need for expertise and saved 24% of technician working time. Application of HCS to antibiotic susceptibility testing of 12 strains demonstrated that it was as efficient as the standard procedure that combines shell vial culture and quantitative PCR.
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high content Screening a reliable system for coxiella burnetii isolation from clinical samples
Journal of Clinical Microbiology, 2020Co-Authors: Rania Francis, Maxime Mioulane, Marion Le Bideau, Mariecharlotte Mati, Pierre-edouard Fournier, Jacques Bou KhalilAbstract:Q fever, caused by Coxiella burnetii, is a worldwide zoonotic disease that may cause severe forms in humans and requires a specific and prolonged antibiotic treatment. Although current serological and molecular detection tools allow a reliable diagnosis of the disease, culture of C. burnetii strains is mandatory to assess their susceptibility to antibiotics and sequence their genome in order to optimize patient management and epidemiological studies. However, cultivating this fastidious microorganism is difficult and restricted to reference centers as it requires biosafety-level 3 laboratories and relies on cell culture performed by experienced technicians. In addition, the culture yield is low, which results in a small number of isolates being available. In this work, we developed a novel high content Screening (HCS) isolation strategy based on optimized high-throughput cell culture and automated microscopic detection of infected cells with specifically-designed algorithms targeting cytopathic effects. This method was more efficient than the shell-vial assay, at the level of time dependency, when applied to both frozen specimens (7 isolates recovered by HCS only, sensitivity 91% vs 78% for shell-vial) and fresh samples (1 additional isolate using HCS, sensitivity 7% vs 5% for shell-vial), for which most strains were recovered more rapidly with the new technique. In addition, detecting positive cultures by an automated microscope reduced the need for expertise and saved 24% of technician working time. Application of HCS to antibiotic susceptibility testing of 12 strains demonstrated that it was as efficient as the standard procedure that combines shell-vial culture and quantitative PCR.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled to high content Screening
Clinical Microbiology and Infection, 2020Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:Objectives A novel coronavirus, SARS-CoV-2, is responsible for the current COVID-19 global pandemic Only a few laboratories routinely isolate the virus, which is because the current co-culture strategy is highly time-consuming and requires working in a biosafety level 3 laboratory This work aimed to develop a new high-throughput isolation strategy using novel technologies for rapid and automated isolation of SARS-CoV-2 Methods We used an automated microscope based on High-Content Screening (HCS), and we applied specific image analysis algorithms targeting cytopathic effects of SARS-CoV-2 on Vero E6 cells A randomized panel of 104 samples, including 72 that tested positive by RT-PCR and 32 that tested negative, were processed with our HCS strategy and were compared to the classical isolation procedure Results The isolation rate was 43% (31/72) with both strategies on RT-PCR-positive samples and was correlated with the initial RNA viral load in the samples, in which we obtained a positivity threshold of 27 Ct Co-culture delays were shorter with the HCS strategy, where 80% (25/31) of the positive samples were recovered by the third day of co-culture, compared to only 26% (8/30) with the classic strategy Moreover, only the HCS strategy allowed us to recover all the positive samples (31 with HCS versus 27 with classic strategy) after 1 week of co-culture Conclusions This system allows the rapid and automated Screening of clinical samples with minimal operator workload, which reduces the risk of contamination, thus paving the way for future applications in clinical microbiology, such as large-scale drug susceptibility testing
Marion Le Bideau - One of the best experts on this subject based on the ideXlab platform.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled to high content Screening
Clinical Microbiology and Infection, 2021Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:Abstract Objectives A novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is responsible for the current coronavirus disease 2019 global pandemic. Only a few laboratories routinely isolate the virus, which is because the current co-culture strategy is highly time-consuming and requires a biosafety level 3 laboratory. This work aimed to develop a new high-throughput isolation strategy using novel technologies for rapid and automated isolation of SARS-CoV-2. Methods We used an automated microscope based on High-Content Screening (HCS), and we applied specific image analysis algorithms targeting cytopathic effects of SARS-CoV-2 on Vero E6 cells. A randomized panel of 104 samples, including 72 that tested positive by RT-PCR and 32 that tested negative, were processed with our HCS strategy and were compared with the classical isolation procedure. Results The isolation rate was 43% (31/72) with both strategies on RT-PCR-positive samples and was correlated with the initial RNA viral load in the samples, in which we obtained a positivity threshold of 27 Ct. Co-culture delays were shorter with the HCS strategy, where 80% (25/31) of the positive samples were recovered by the third day of co-culture, compared with only 26% (8/30) with the classic strategy. Moreover, only the HCS strategy allowed us to recover all the positive samples (31 with HCS versus 27 with classic strategy) after 1 week of co-culture. Conclusions This system allows the rapid and automated Screening of clinical samples with minimal operator workload, which reduces the risk of contamination and paves the way for future applications in clinical microbiology, such as large-scale drug susceptibility testing.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled with high content Screening
bioRxiv, 2020Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:SARS-CoV-2, a novel coronavirus infecting humans, is responsible for the current COVID-19 global pandemic. If several strains could be isolated worldwide, especially for in-vitro drug susceptibility testing and vaccine development, few laboratories routinely isolate SARS-CoV-2. This is due to the fact that the current co-culture strategy is highly time consuming and requires working in a biosafety level 3 laboratory. In this work, we present a new strategy based on high content Screening automated microscopy (HCS) allowing large scale isolation of SARS-CoV-2 from clinical samples in 1 week. A randomized panel of 104 samples, including 72 tested positive by RT-PCR and 32 tested negative, were processed with our HCS procedure and were compared to the classical isolation procedure. Isolation rate was 43% with both strategies on RT-PCR positive samples, and was correlated with the initial RNA viral load in the samples, where we obtained a positivity threshold of 27Ct. Co-culture delays were shorter with HCS strategy, where 80% of the positive samples were recovered by the third day of co-culture, as compared to only 25% with the classic strategy. Moreover, only the HCS strategy allowed us to recover all the positive elements after 1 week of co-culture. This system allows rapid and automated Screening of clinical samples with minimal operator intervention, thus accelerating the work rate and minimizing the risk of contamination of the laboratory personnel.
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high content Screening a reliable system for coxiella burnetii isolation from clinical samples
Journal of Clinical Microbiology, 2020Co-Authors: Rania Francis, Maxime Mioulane, Marion Le Bideau, Mariecharlotte Mati, Jacques Yaacoub Bou Khalil, Pierre-edouard Fournier, Bernard La ScolaAbstract:ABSTRACT Q fever, caused by Coxiella burnetii, is a worldwide zoonotic disease that may cause severe forms in humans and requires a specific and prolonged antibiotic treatment. Although current serological and molecular detection tools allow a reliable diagnosis of the disease, culture of C. burnetii strains is mandatory to assess their susceptibility to antibiotics and sequence their genome in order to optimize patient management and epidemiological studies. However, cultivating this fastidious microorganism is difficult and restricted to reference centers, as it requires biosafety level 3 laboratories and relies on cell culture performed by experienced technicians. In addition, the culture yield is low, which results in a small number of isolates being available. In this work, we developed a novel High-Content Screening (HCS) isolation strategy based on optimized high-throughput cell culture and automated microscopic detection of infected cells with specifically designed algorithms targeting cytopathic effects. This method was more efficient than the shell vial assay, at the level of time dependency, when applied to both frozen specimens (7 isolates recovered by HCS only, sensitivity 91% versus 78% for shell vial) and fresh samples (1 additional isolate using HCS, sensitivity 7% versus 5% for shell vial), for which most strains were recovered more rapidly with the new technique. In addition, detecting positive cultures by an automated microscope reduced the need for expertise and saved 24% of technician working time. Application of HCS to antibiotic susceptibility testing of 12 strains demonstrated that it was as efficient as the standard procedure that combines shell vial culture and quantitative PCR.
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high content Screening a reliable system for coxiella burnetii isolation from clinical samples
Journal of Clinical Microbiology, 2020Co-Authors: Rania Francis, Maxime Mioulane, Marion Le Bideau, Mariecharlotte Mati, Pierre-edouard Fournier, Jacques Bou KhalilAbstract:Q fever, caused by Coxiella burnetii, is a worldwide zoonotic disease that may cause severe forms in humans and requires a specific and prolonged antibiotic treatment. Although current serological and molecular detection tools allow a reliable diagnosis of the disease, culture of C. burnetii strains is mandatory to assess their susceptibility to antibiotics and sequence their genome in order to optimize patient management and epidemiological studies. However, cultivating this fastidious microorganism is difficult and restricted to reference centers as it requires biosafety-level 3 laboratories and relies on cell culture performed by experienced technicians. In addition, the culture yield is low, which results in a small number of isolates being available. In this work, we developed a novel high content Screening (HCS) isolation strategy based on optimized high-throughput cell culture and automated microscopic detection of infected cells with specifically-designed algorithms targeting cytopathic effects. This method was more efficient than the shell-vial assay, at the level of time dependency, when applied to both frozen specimens (7 isolates recovered by HCS only, sensitivity 91% vs 78% for shell-vial) and fresh samples (1 additional isolate using HCS, sensitivity 7% vs 5% for shell-vial), for which most strains were recovered more rapidly with the new technique. In addition, detecting positive cultures by an automated microscope reduced the need for expertise and saved 24% of technician working time. Application of HCS to antibiotic susceptibility testing of 12 strains demonstrated that it was as efficient as the standard procedure that combines shell-vial culture and quantitative PCR.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled to high content Screening
Clinical Microbiology and Infection, 2020Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:Objectives A novel coronavirus, SARS-CoV-2, is responsible for the current COVID-19 global pandemic Only a few laboratories routinely isolate the virus, which is because the current co-culture strategy is highly time-consuming and requires working in a biosafety level 3 laboratory This work aimed to develop a new high-throughput isolation strategy using novel technologies for rapid and automated isolation of SARS-CoV-2 Methods We used an automated microscope based on High-Content Screening (HCS), and we applied specific image analysis algorithms targeting cytopathic effects of SARS-CoV-2 on Vero E6 cells A randomized panel of 104 samples, including 72 that tested positive by RT-PCR and 32 that tested negative, were processed with our HCS strategy and were compared to the classical isolation procedure Results The isolation rate was 43% (31/72) with both strategies on RT-PCR-positive samples and was correlated with the initial RNA viral load in the samples, in which we obtained a positivity threshold of 27 Ct Co-culture delays were shorter with the HCS strategy, where 80% (25/31) of the positive samples were recovered by the third day of co-culture, compared to only 26% (8/30) with the classic strategy Moreover, only the HCS strategy allowed us to recover all the positive samples (31 with HCS versus 27 with classic strategy) after 1 week of co-culture Conclusions This system allows the rapid and automated Screening of clinical samples with minimal operator workload, which reduces the risk of contamination, thus paving the way for future applications in clinical microbiology, such as large-scale drug susceptibility testing
Jacques Yaacoub Bou Khalil - One of the best experts on this subject based on the ideXlab platform.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled to high content Screening
Clinical Microbiology and Infection, 2021Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:Abstract Objectives A novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is responsible for the current coronavirus disease 2019 global pandemic. Only a few laboratories routinely isolate the virus, which is because the current co-culture strategy is highly time-consuming and requires a biosafety level 3 laboratory. This work aimed to develop a new high-throughput isolation strategy using novel technologies for rapid and automated isolation of SARS-CoV-2. Methods We used an automated microscope based on High-Content Screening (HCS), and we applied specific image analysis algorithms targeting cytopathic effects of SARS-CoV-2 on Vero E6 cells. A randomized panel of 104 samples, including 72 that tested positive by RT-PCR and 32 that tested negative, were processed with our HCS strategy and were compared with the classical isolation procedure. Results The isolation rate was 43% (31/72) with both strategies on RT-PCR-positive samples and was correlated with the initial RNA viral load in the samples, in which we obtained a positivity threshold of 27 Ct. Co-culture delays were shorter with the HCS strategy, where 80% (25/31) of the positive samples were recovered by the third day of co-culture, compared with only 26% (8/30) with the classic strategy. Moreover, only the HCS strategy allowed us to recover all the positive samples (31 with HCS versus 27 with classic strategy) after 1 week of co-culture. Conclusions This system allows the rapid and automated Screening of clinical samples with minimal operator workload, which reduces the risk of contamination and paves the way for future applications in clinical microbiology, such as large-scale drug susceptibility testing.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled with high content Screening
bioRxiv, 2020Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:SARS-CoV-2, a novel coronavirus infecting humans, is responsible for the current COVID-19 global pandemic. If several strains could be isolated worldwide, especially for in-vitro drug susceptibility testing and vaccine development, few laboratories routinely isolate SARS-CoV-2. This is due to the fact that the current co-culture strategy is highly time consuming and requires working in a biosafety level 3 laboratory. In this work, we present a new strategy based on high content Screening automated microscopy (HCS) allowing large scale isolation of SARS-CoV-2 from clinical samples in 1 week. A randomized panel of 104 samples, including 72 tested positive by RT-PCR and 32 tested negative, were processed with our HCS procedure and were compared to the classical isolation procedure. Isolation rate was 43% with both strategies on RT-PCR positive samples, and was correlated with the initial RNA viral load in the samples, where we obtained a positivity threshold of 27Ct. Co-culture delays were shorter with HCS strategy, where 80% of the positive samples were recovered by the third day of co-culture, as compared to only 25% with the classic strategy. Moreover, only the HCS strategy allowed us to recover all the positive elements after 1 week of co-culture. This system allows rapid and automated Screening of clinical samples with minimal operator intervention, thus accelerating the work rate and minimizing the risk of contamination of the laboratory personnel.
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high content Screening a reliable system for coxiella burnetii isolation from clinical samples
Journal of Clinical Microbiology, 2020Co-Authors: Rania Francis, Maxime Mioulane, Marion Le Bideau, Mariecharlotte Mati, Jacques Yaacoub Bou Khalil, Pierre-edouard Fournier, Bernard La ScolaAbstract:ABSTRACT Q fever, caused by Coxiella burnetii, is a worldwide zoonotic disease that may cause severe forms in humans and requires a specific and prolonged antibiotic treatment. Although current serological and molecular detection tools allow a reliable diagnosis of the disease, culture of C. burnetii strains is mandatory to assess their susceptibility to antibiotics and sequence their genome in order to optimize patient management and epidemiological studies. However, cultivating this fastidious microorganism is difficult and restricted to reference centers, as it requires biosafety level 3 laboratories and relies on cell culture performed by experienced technicians. In addition, the culture yield is low, which results in a small number of isolates being available. In this work, we developed a novel High-Content Screening (HCS) isolation strategy based on optimized high-throughput cell culture and automated microscopic detection of infected cells with specifically designed algorithms targeting cytopathic effects. This method was more efficient than the shell vial assay, at the level of time dependency, when applied to both frozen specimens (7 isolates recovered by HCS only, sensitivity 91% versus 78% for shell vial) and fresh samples (1 additional isolate using HCS, sensitivity 7% versus 5% for shell vial), for which most strains were recovered more rapidly with the new technique. In addition, detecting positive cultures by an automated microscope reduced the need for expertise and saved 24% of technician working time. Application of HCS to antibiotic susceptibility testing of 12 strains demonstrated that it was as efficient as the standard procedure that combines shell vial culture and quantitative PCR.
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high speed large scale automated isolation of sars cov 2 from clinical samples using miniaturized co culture coupled to high content Screening
Clinical Microbiology and Infection, 2020Co-Authors: Rania Francis, Marion Le Bideau, Jacques Yaacoub Bou Khalil, Priscilla Jardot, Clio Grimaldier, Bernard La ScolaAbstract:Objectives A novel coronavirus, SARS-CoV-2, is responsible for the current COVID-19 global pandemic Only a few laboratories routinely isolate the virus, which is because the current co-culture strategy is highly time-consuming and requires working in a biosafety level 3 laboratory This work aimed to develop a new high-throughput isolation strategy using novel technologies for rapid and automated isolation of SARS-CoV-2 Methods We used an automated microscope based on High-Content Screening (HCS), and we applied specific image analysis algorithms targeting cytopathic effects of SARS-CoV-2 on Vero E6 cells A randomized panel of 104 samples, including 72 that tested positive by RT-PCR and 32 that tested negative, were processed with our HCS strategy and were compared to the classical isolation procedure Results The isolation rate was 43% (31/72) with both strategies on RT-PCR-positive samples and was correlated with the initial RNA viral load in the samples, in which we obtained a positivity threshold of 27 Ct Co-culture delays were shorter with the HCS strategy, where 80% (25/31) of the positive samples were recovered by the third day of co-culture, compared to only 26% (8/30) with the classic strategy Moreover, only the HCS strategy allowed us to recover all the positive samples (31 with HCS versus 27 with classic strategy) after 1 week of co-culture Conclusions This system allows the rapid and automated Screening of clinical samples with minimal operator workload, which reduces the risk of contamination, thus paving the way for future applications in clinical microbiology, such as large-scale drug susceptibility testing
Lansing D Taylor - One of the best experts on this subject based on the ideXlab platform.
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characterization and optimization of a novel protein protein interaction biosensor high content Screening assay to identify disruptors of the interactions between p53 and hdm2
Assay and Drug Development Technologies, 2010Co-Authors: Drew D Dudgeon, Kenneth A Giuliano, Lansing D Taylor, Patricia A. Johnston, Sunita Shinde, Tong Ying Shun, John S Lazo, Christopher J Strock, Paul A JohnstonAbstract:Abstract We present here the characterization and optimization of a novel imaging-based positional biosensor High-Content Screening (HCS) assay to identify disruptors of p53-hDM2 protein–protein interactions (PPIs). The chimeric proteins of the biosensor incorporated the N-terminal PPI domains of p53 and hDM2, protein targeting sequences (nuclear localization and nuclear export sequence), and fluorescent reporters, which when expressed in cells could be used to monitor p53-hDM2 PPIs through changes in the subcellular localization of the hDM2 component of the biosensor. Coinfection with the recombinant adenovirus biosensors was used to express the NH-terminal domains of p53 and hDM2, fused to green fluorescent protein and red fluorescent protein, respectively, in U-2 OS cells. We validated the p53-hDM2 PPI biosensor (PPIB) HCS assay with Nutlin-3, a compound that occupies the hydrophobic pocket on the surface of the N-terminus of hDM2 and blocks the binding interactions with the N-terminus of p53. Nutlin-3...
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[31] systems cell biology 601 [31] Systems Cell Biology Based on High‐Content Screening
2008Co-Authors: A. Giuliano, Albert Gough, Patricia A. Johnston, Lansing D TaylorAbstract:A new discipline of biology has emerged since 2004, which we call ‘‘systems cell biology’ ’ (SCB). Systems cell biology is the study of the living cell, the basic unit of life, an integrated and interacting network of genes, proteins, and myriad metabolic reactions that give rise to function. SCB takes advantage of high‐content Screening platforms, but delivers more detailed profiles of cellular systemic function, including the application of advanced reagents and informatics tools to sophisticated cellular models. Therefore, an SCB profile is a cellular systemic response as measured by a panel of reagents that quantify a specific set of biomarkers. Background Until recently, the focus in drug discovery and basic biomedical research has been on simplifying the complexity of the living human organism to individual genes, single metabolic pathways, single proteins, and one potential modulating molecule, such as a small chemical compound or bioproducts t
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high content Screening applied to large scale cell biology
Trends in Biotechnology, 2004Co-Authors: Vivek C Abraham, Lansing D Taylor, Jeffrey R HaskinsAbstract:Abstract High Content Screening (HCS) has been useful as an early drug-discovery platform for defining the functions of genes, proteins and other biomolecules in normal and abnormal cellular functions. The same concept of integrating and automating cell-sample preparation, fluorescent labeling, image acquisition, image processing, image analysis, information management and knowledge mining is now beginning to have an impact in basic biomedical research. These integrated tools allow an increase in the scale of cell biological investigations to large-scale cell biology and their value is demonstrated by the rapid performance of a kinetic, multiparameter cytotoxicity assay enabling generation of 10 8 individual physiological cellular measurements in less than one day.
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streamlining the drug discovery process by integrating miniaturization high throughput Screening high content Screening and automation on the cellchip system
Biomedical Microdevices, 1999Co-Authors: Ravi Kapur, Kenneth A Giuliano, Martha Campana, Terri Adams, Keith Olson, David R Jung, Milan Mrksich, Chandrasekaran Vasudevan, Lansing D TaylorAbstract:A major bottleneck to the early stages of drug discovery is the absence of integration of high throughput Screening (HTS) with smarter assays that screen “hits” from HTS to identify leads (High content Screening, HCS). We propose a solution using novel fluorescent engineered protein biosensors integrated into a miniaturized live-cell-based Screening platform (CellChip™ System) that markedly shortens the early drug discovery process. Microarrays of selectively localized living cells, containing engineered fluorescent biosensors, serve to integrate HTS and HCS onto a single platform. HTS “hits” are identified using one biosensor while reading the whole chip array of cells. The high-biological content information is then obtained from probing target activity at inter-cellular, sub-cellular and molecular levels in the “hit” wells. HCS assays yield temporal-spatial dynamic maps of the drug-target interaction within each living cell. We predict that a new platform incorporating HTS and HCS assays that are automated, miniaturized, and information-rich will dramatically improve the decision making process in the pharmaceutical industry and optimize lead compounds during the early part of the drug discovery process. There is an opportunity to establish a new paradigm for drug discovery based on integration of fluorescence technology, micropatterning of living cells, automated optical detection and data analysis, and a new generation of knowledge building bioinformatics approaches. The technology will have an expansive impact spanning the fields of drug discovery, biomedical research, environmental monitoring, life sciences, and clinical diagnostics. The integrated CellChip™ Platform with miniaturized tissue-specific microarrayed cells capable of providing inter-cellular and sub-cellular spatio-temporal information in response to drug-cell, toxin-cell, or pathogen-cell interactions will serve to enhance the decision making process in drug discovery, toxicology, and clinical diagnostics.
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high content Screening a new approach to easing key bottlenecks in the drug discovery process
Journal of Biomolecular Screening, 1997Co-Authors: Kenneth A Giuliano, Robbin Debiasio, Terry R Dunlay, Albert Gough, Joanne M Volosky, Joseph Zock, George N Pavlakis, Lansing D TaylorAbstract:Recent improvements in target discovery and high throughput Screening (HTS) have increased the pressure at key points along the drug discovery pipeline. High-Content Screening (HCS) was developed t...