Comparative sensitivity of four different cell lines for the isolation of Coxiella burnetii

Lockhart, Michelle G.; Islam, Aminul; Fenwick, Stan; Graves, Stephen; Stenos, John

doi:10.1111/j.1574-6968.2012.02617.x

Cited by 10 publications

(7 citation statements)

References 9 publications

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“…The introduction of a diverse panel of cell lines for the isolation of C. burnetii could increase the efficiency of the system by making it possible to isolate strains with different affinities for different cell lines. Such variations in affinity for C. burnetii have already been described in the literature (26,27). This system would also be applicable for tropism and virulence assessment.…”

Section: Discussionsupporting

confidence: 53%

High-Content Screening, a Reliable System for Coxiella burnetii Isolation from Clinical Samples

Francis

Mioulane²,

Bideau

et al. 2020

J Clin Microbiol

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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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Section: Discussionsupporting

confidence: 53%

High-Content Screening, a Reliable System for Coxiella burnetii Isolation from Clinical Samples

Francis

Mioulane²,

Bideau

et al. 2020

J Clin Microbiol

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“…Gimenez or immunofluorescence staining is used for detection of the bacterium inside cells. Lockhart et al have compared four different cell lines for the isolation of C. burnetii using two different isolates, the Henzerling and Arandale strains (364). For the Henzerling strain, DH82 cells were the most sensitive, while for the Arandale isolate, Vero cells showed the highest sensitivity.…”

Section: Culturementioning

confidence: 99%

From Q Fever to Coxiella burnetii Infection: a Paradigm Change

et al. 2017

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Coxiella burnetii is the agent of Q fever, or "query fever," a zoonosis first described in Australia in 1937. Since this first description, knowledge about this pathogen and its associated infections has increased dramatically. We review here all the progress made over the last 20 years on this topic. C. burnetii is classically a strict intracellular, Gram-negative bacterium. However, a major step in the characterization of this pathogen was achieved by the establishment of its axenic culture. C. burnetii infects a wide range of animals, from arthropods to humans. The genetic determinants of virulence are now better known, thanks to the achievement of determining the genome sequences of several strains of this species and comparative genomic analyses. Q fever can be found worldwide, but the epidemiological features of this disease vary according to the geographic area considered, including situations where it is endemic or hyperendemic, and the occurrence of large epidemic outbreaks. In recent years, a major breakthrough in the understanding of the natural history of human infection with C. burnetii was the breaking of the old dichotomy between "acute" and "chronic" Q fever. The clinical presentation of C. burnetii infection depends on both the virulence of the infecting C. burnetii strain and specific risks factors in the infected patient. Moreover, no persistent infection can exist without a focus of infection. This paradigm change should allow better diagnosis and management of primary infection and long-term complications in patients with C. burnetii infection.

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“…The introduction of a panel of cell lines for the isolation of C. burnetii may increase the system’s efficiency to isolate more strains having different susceptibilities. Previous studies have already described the variation of the susceptibility of C. burnetii to different cell lines, as well as strain dependent susceptibility (26),(27). This system would also be applicable for tropism and virulence assessment.…”

Section: Discussionmentioning

confidence: 99%

High Content Screening, a reliable system forCoxiella burnetiiisolation from clinical samples

Francis

Mioulane²,

Bideau

et al. 2019

Preprint

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24Q fever, caused by Coxiella burnetii, is a worldwide zoonotic disease that may cause severe 25 forms in humans and requires a specific and prolonged antibiotic treatment. Although the current 26 serological and molecular detection tools enable a reliable diagnosis of the disease, culture of C. 27 burnetii strains is mandatory to evaluate their antibiotic susceptibility and sequence their genome 28 in order to optimize patient management and epidemiological studies. However, cultivating this 29 fastidious microorganism is difficult and restricted to reference centers as it requires biosafety-30 level 3 laboratories and relies on cell culture performed by experienced technicians. In addition, 31 the culture yield is low, which results in a small number of isolates being available. In this work, 32 we developed a novel high content screening (HCS) isolation strategy based on optimized high-33 throughput cell culture and automated microscopic detection of infected cells with specifically-34 designed algorithms targeting cytopathic effects. This method was more efficient than the shell-35 vial assay when applied to both frozen specimens (7 isolates recovered by HCS only, sensitivity 36 91% vs 78% for shell-vial) and fresh samples (1 additional isolate using HCS, sensitivity 7% vs 37 5% for shell-vial). In addition, detecting positive cultures by an automated microscope reduced 38 the need for expertise and saved 24% of technician working time. Application of HCS to 39 antibiotic susceptibility testing of 12 strains demonstrated that it was as efficient as the standard 40 procedure that combines shell-vial culture and quantitative PCR. Overall, this high-throughput 41 HCS system paves the way to the development of improved cell culture isolation of human 42 viruses.43

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Comparative sensitivity of four different cell lines for the isolation of Coxiella burnetii

Cited by 10 publications

References 9 publications

High-Content Screening, a Reliable System for Coxiella burnetii Isolation from Clinical Samples

High-Content Screening, a Reliable System for Coxiella burnetii Isolation from Clinical Samples

From Q Fever to Coxiella burnetii Infection: a Paradigm Change

High Content Screening, a reliable system forCoxiella burnetiiisolation from clinical samples

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