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  1. Article: Extraction-Free Detection of SARS-CoV-2 Viral RNA Using LumiraDx's RNA Star Complete Assay from Clinical Nasal Swabs Stored in a Novel Collection and Transport Medium.

    Daum, Luke T / Rodriguez, John D / Ward, Susan R / Chambers, James P

    Diagnostics (Basel, Switzerland)

    2023  Volume 13, Issue 18

    Abstract: Background: ...

    Abstract Background:
    Language English
    Publishing date 2023-09-21
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2662336-5
    ISSN 2075-4418
    ISSN 2075-4418
    DOI 10.3390/diagnostics13183010
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  2. Article: Rapid and Safe Detection of SARS-CoV-2 and Influenza Virus RNA Using Onsite Quantitative PCR Diagnostic Testing From Clinical Specimens Collected in Molecular Transport Medium.

    Daum, Luke T / Fischer, Gerald W

    The journal of applied laboratory medicine

    2021  Volume 6, Issue 6, Page(s) 1409–1416

    Abstract: Background: The ability to rapidly detect severe accurate respiratory syndrome coronavirus virus-2 (SARS-CoV-2) and influenza virus infection is vital for patient care due to overlap in clinical symptoms. Roche's cobas® Liat® SARS-CoV-2 & Influenza A/B ... ...

    Abstract Background: The ability to rapidly detect severe accurate respiratory syndrome coronavirus virus-2 (SARS-CoV-2) and influenza virus infection is vital for patient care due to overlap in clinical symptoms. Roche's cobas® Liat® SARS-CoV-2 & Influenza A/B Nucleic Acid Test used on the cobas Liat was granted approval under the Food and Drug's Emergency Use Authorization for nasopharyngeal (NP) and nasal swabs collected in viral/universal transport medium (VTM/UTM). However, there is a critical need for media that inactivates the virus, especially when specimens are collected in decentralized settings. This study aimed to investigate the use of PrimeStore Molecular Transport Medium® (PS-MTM®), designed to inactivate/kill and stabilize RNA/DNA for ambient transport and preprocessing of collected samples.
    Methods: A limit of detection (LOD) using serially diluted SARS-CoV-2 RNA in PS-MTM and routine UTM was established using standard quantitative PCR (qPCR). Additionally, a clinical panel of NP and oral swabs collected in PS-MTM during the 2020 coronavirus disease 2019 pandemic were evaluated on the cobas Liat and compared to "gold standard" qPCR on an ABI-7500 instrument.
    Results: SARS-CoV-2 RNA LOD using standard qPCR was equivalent on the cobas Liat instrument. cobas Liat detection from oral/NP swabs in PS-MTM media exhibited equivalent positive percent agreement (100%) and negative percent agreement (96.4%).
    Conclusion: PS-MTM and the Roche cobas Liat are compatible and complimentary devices for respiratory specimen collection and rapid disease detection, respectively. PS-MTM is equivalent to standard VTM/UTM with the added benefit of safe, noninfectious sample processing for near-patient testing.
    MeSH term(s) COVID-19 ; Humans ; Orthomyxoviridae ; RNA, Viral/genetics ; Real-Time Polymerase Chain Reaction ; SARS-CoV-2 ; Specimen Handling
    Chemical Substances RNA, Viral
    Language English
    Publishing date 2021-06-21
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 2576-9456
    ISSN 2576-9456
    DOI 10.1093/jalm/jfab073
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  3. Article ; Online: Extraction-Free Detection of SARS-CoV-2 Viral RNA Using LumiraDx’s RNA Star Complete Assay from Clinical Nasal Swabs Stored in a Novel Collection and Transport Medium

    Luke T. Daum / John D. Rodriguez / Susan R. Ward / James P. Chambers

    Diagnostics, Vol 13, Iss 3010, p

    2023  Volume 3010

    Abstract: ... qPCR cycle threshold (C T ) values for vRNA in XF and UTM were observed to be equivalent. An LOD ... negative when tested via RSC; however, all three samples had C T values ≥ 36.4. Conclusions: XF is ...

    Abstract Background: The rapid detection of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is vital for patient care. The LumiraDx™ SARS-CoV-2 RNA Star Complete (RSC) is an Emergency Use Authorization -recognized molecular test using nasal/nasopharyngeal swabs immersed in a viral/universal transport medium (VTM/UTM). However, there is a critical need for an alternative medium for point-of-care testing (POCT). This study aimed to investigate Xtract-Free (XF), a novel collection medium for transport and direct (extraction-free) use with nucleic acid tests. Methods: Using serially diluted SARS-CoV-2 viral RNA (vRNA) in a routine UTM and XF, a limit of detection (LOD) was established via an RSC test and a quantitative reverse transcription PCR (RT-qPCR). Additionally, the results obtained from a panel of 108 clinical “car-side” nasal swabs collected in XF during the coronavirus pandemic and assessed using the ”gold-standard” RT-qPCR assay were compared to Lumira’s RSC assay. Results: The average replicate RT-qPCR cycle threshold (C T ) values for vRNA in XF and UTM were observed to be equivalent. An LOD for which five out of five replicates were detected using XF or VTM was approximately 2000 copies/mL. The nasal swabs collected in XF exhibited 93.9% positive percent agreement (sensitivity) and 100% negative percent agreement (specificity) compared to the RT-qPCR. Three specimens tested positive via an RT-qPCR were negative when tested via RSC; however, all three samples had C T values ≥ 36.4. Conclusions: XF is equivalent to VTM/UTM and is compatible for use with the RSC test. Furthermore, XF can be used directly with RT-qPCRs and rapid antigen testing without the requirement for separate nucleic acid extraction (an extraction-free process), making it ideal for cost-effective high-throughput and decentralized respiratory testing. Impact Statement: This study is the first to evaluate LumiraDx’s SARS-CoV-2 RNA Star Complete assay in concert with Xtract-Free (XF), a novel collection medium containing a ...
    Keywords Xtract-Free medium ; sample collection ; qPCR ; diagnostics ; COVID-19 ; SARS-CoV-2 ; Medicine (General) ; R5-920
    Subject code 610
    Language English
    Publishing date 2023-09-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article: Conserved Influenza Hemagglutinin, Neuraminidase and Matrix Peptides Adjuvanted with ALFQ Induce Broadly Neutralizing Antibodies.

    Sei, Clara J / Rao, Mangala / Schuman, Richard F / Daum, Luke T / Matyas, Gary R / Rikhi, Nimisha / Muema, Kevin / Anderson, Alexander / Jobe, Ousman / Kroscher, Kellie A / Alving, Carl R / Fischer, Gerald W

    Vaccines

    2021  Volume 9, Issue 7

    Abstract: A universal influenza candidate vaccine that targets multiple conserved influenza virus epitopes from hemagglutinin (HA), neuraminidase (NA) and matrix (M2e) proteins was combined with the potent Army liposomal adjuvant (ALFQ) to promote induction of ... ...

    Abstract A universal influenza candidate vaccine that targets multiple conserved influenza virus epitopes from hemagglutinin (HA), neuraminidase (NA) and matrix (M2e) proteins was combined with the potent Army liposomal adjuvant (ALFQ) to promote induction of broad immunity to seasonal and pandemic influenza strains. The unconjugated and CRM-conjugated composite peptides formulated with ALFQ were highly immunogenic and induced both humoral and cellular immune responses in mice. Broadly reactive serum antibodies were induced across various IgG isotypes. Mice immunized with the unconjugated composite peptide developed antibody responses earlier than mice immunized with conjugated peptides, and the IgG antibodies were broadly reactive and neutralizing across Groups 1 and 2 influenza viruses. Multi-epitope unconjugated influenza composite peptides formulated with ALFQ provide a novel strategy for the development of a universal influenza vaccine. These synthetic peptide vaccines avoid the pitfalls of egg-produced influenza vaccines and production can be scaled up rapidly and economically.
    Language English
    Publishing date 2021-06-25
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2703319-3
    ISSN 2076-393X
    ISSN 2076-393X
    DOI 10.3390/vaccines9070698
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  5. Article ; Online: Conserved Influenza Hemagglutinin, Neuraminidase and Matrix Peptides Adjuvanted with ALFQ Induce Broadly Neutralizing Antibodies

    Clara J. Sei / Mangala Rao / Richard F. Schuman / Luke T. Daum / Gary R. Matyas / Nimisha Rikhi / Kevin Muema / Alexander Anderson / Ousman Jobe / Kellie A. Kroscher / Carl R. Alving / Gerald W. Fischer

    Vaccines, Vol 9, Iss 698, p

    2021  Volume 698

    Abstract: A universal influenza candidate vaccine that targets multiple conserved influenza virus epitopes from hemagglutinin (HA), neuraminidase (NA) and matrix (M2e) proteins was combined with the potent Army liposomal adjuvant (ALFQ) to promote induction of ... ...

    Abstract A universal influenza candidate vaccine that targets multiple conserved influenza virus epitopes from hemagglutinin (HA), neuraminidase (NA) and matrix (M2e) proteins was combined with the potent Army liposomal adjuvant (ALFQ) to promote induction of broad immunity to seasonal and pandemic influenza strains. The unconjugated and CRM-conjugated composite peptides formulated with ALFQ were highly immunogenic and induced both humoral and cellular immune responses in mice. Broadly reactive serum antibodies were induced across various IgG isotypes. Mice immunized with the unconjugated composite peptide developed antibody responses earlier than mice immunized with conjugated peptides, and the IgG antibodies were broadly reactive and neutralizing across Groups 1 and 2 influenza viruses. Multi-epitope unconjugated influenza composite peptides formulated with ALFQ provide a novel strategy for the development of a universal influenza vaccine. These synthetic peptide vaccines avoid the pitfalls of egg-produced influenza vaccines and production can be scaled up rapidly and economically.
    Keywords peptides ; influenza ; ALFQ ; liposomes ; QS21 (also known as QS-21) ; universal vaccine ; Medicine ; R
    Language English
    Publishing date 2021-06-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article: Erratum to "Opsonic monoclonal antibodies enhance phagocytic killing activity and clearance of

    Sei, Clara J / Shey, Bong-Akee / Schuman, Richard F / Rikhi, Nimisha / Muema, Kevin / Rodriguez, John D / Daum, Luke T / Fourie, P Bernard / Fischer, Gerald W

    Heliyon

    2019  Volume 5, Issue 10, Page(s) e02627

    Abstract: This corrects the article DOI: 10.1016/j.heliyon.2019.e02260.]. ...

    Abstract [This corrects the article DOI: 10.1016/j.heliyon.2019.e02260.].
    Language English
    Publishing date 2019-11-01
    Publishing country England
    Document type Published Erratum
    ZDB-ID 2835763-2
    ISSN 2405-8440
    ISSN 2405-8440
    DOI 10.1016/j.heliyon.2019.e02627
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  7. Article: Opsonic monoclonal antibodies enhance phagocytic killing activity and clearance of

    Sei, Clara J / Shey, Bong-Akee / Schuman, Richard F / Rikhi, Nimisha / Muema, Kevin / Rodriguez, John D / Daum, Luke T / Fourie, P Bernard / Fischer, Gerald W

    Heliyon

    2019  Volume 5, Issue 9, Page(s) e02260

    Abstract: Background: Patients with impaired immunity often have rapid progression of tuberculosis (TB) which can lead to highly lethal : Methods: BALB/c mice were immunized with ethanol-killed MTB (EK-MTB) and MABs were produced and screened by ELISA for ... ...

    Abstract Background: Patients with impaired immunity often have rapid progression of tuberculosis (TB) which can lead to highly lethal
    Methods: BALB/c mice were immunized with ethanol-killed MTB (EK-MTB) and MABs were produced and screened by ELISA for binding to killed and live
    Results: Two MABs (GG9 and JG7) bound to killed and live SMEG and MTB (susceptible and resistant), and promoted OPKA with live MTB. MAB JG7 significantly enhanced OPKA of MTB. Both MABs significantly enhanced clearance of killed MTB from murine blood at 4 and 24 h as measured by qPCR. These opsonic MABs bound to PGN, a major cell wall constituent.
    Conclusions: Anti-MTB MABs that promote bactericidal phagocytic activity of MTB and enhance clearance of killed MTB from the blood, may offer an immunotherapeutic approach for treatment of MTB bacteremia or sepsis, and augment treatment of multi-drug resistant (MDR) or extensively drug resistant (XDR) TB.
    Language English
    Publishing date 2019-09-06
    Publishing country England
    Document type Journal Article
    ZDB-ID 2835763-2
    ISSN 2405-8440
    ISSN 2405-8440
    DOI 10.1016/j.heliyon.2019.e02260
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  8. Article ; Online: Erratum to “Opsonic monoclonal antibodies enhance phagocytic killing activity and clearance of Mycobacterium tuberculosis from blood in a quantitative qPCR mouse model” [Heliyon 5 (9), (September 2019), e02260]

    Clara J. Sei / Bong-Akee Shey / Richard F. Schuman / Nimisha Rikhi / Kevin Muema / John D. Rodriguez / Luke T. Daum / P Bernard Fourie / Gerald W. Fischer

    Heliyon, Vol 5, Iss 10, Pp e02627- (2019)

    2019  

    Keywords Science (General) ; Q1-390 ; Social sciences (General) ; H1-99
    Language English
    Publishing date 2019-10-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  9. Article ; Online: Multi- and Extensively Drug Resistant Mycobacterium tuberculosis in South Africa: a Molecular Analysis of Historical Isolates.

    Maningi, Nontuthuko E / Daum, Luke T / Rodriguez, John D / Said, Halima M / Peters, Remco P H / Sekyere, John Osei / Fischer, Gerald W / Chambers, James P / Fourie, P Bernard

    Journal of clinical microbiology

    2018  Volume 56, Issue 5

    Abstract: Modern advances in genomics provide an opportunity to reinterpret historical bacterial culture collections. In this study, genotypic antibiotic resistance profiles ... ...

    Abstract Modern advances in genomics provide an opportunity to reinterpret historical bacterial culture collections. In this study, genotypic antibiotic resistance profiles of
    MeSH term(s) Adult ; Antitubercular Agents/pharmacology ; DNA, Bacterial/genetics ; DNA, Bacterial/isolation & purification ; Drug Resistance, Multiple, Bacterial ; Female ; Genes, Bacterial ; Genotype ; Humans ; Male ; Mutation ; Mycobacterium tuberculosis/drug effects ; Mycobacterium tuberculosis/genetics ; Mycobacterium tuberculosis/isolation & purification ; Nucleic Acid Hybridization ; Sequence Analysis, DNA ; South Africa
    Chemical Substances Antitubercular Agents ; DNA, Bacterial
    Language English
    Publishing date 2018-04-25
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 390499-4
    ISSN 1098-660X ; 0095-1137
    ISSN (online) 1098-660X
    ISSN 0095-1137
    DOI 10.1128/JCM.01214-17
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Order via subito

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  10. Article ; Online: Opsonic monoclonal antibodies enhance phagocytic killing activity and clearance of Mycobacterium tuberculosis from blood in a quantitative qPCR mouse model

    Sei, Clara J. / Shey, Bong-Akee / Schuman, Richard F. / Rikhi, Nimisha / Muema, Kevin / Rodriguez, John D. / Daum, Luke T. / Fourie, P. Bernard / Fischer, Gerald W.

    Heliyon. 2019 Sept., v. 5, no. 9 p.e02260-

    2019  

    Abstract: Patients with impaired immunity often have rapid progression of tuberculosis (TB) which can lead to highly lethal Mycobacterium tuberculosis (MTB) sepsis. Opsonic monoclonal antibodies (MABs) directed against MTB that enhance phagocytic killing activity ... ...

    Abstract Patients with impaired immunity often have rapid progression of tuberculosis (TB) which can lead to highly lethal Mycobacterium tuberculosis (MTB) sepsis. Opsonic monoclonal antibodies (MABs) directed against MTB that enhance phagocytic killing activity and clearance of MTB from blood may be useful to enhance TB immunity. BALB/c mice were immunized with ethanol-killed MTB (EK-MTB) and MABs were produced and screened by ELISA for binding to killed and live Mycobacterium smegmatis (SMEG) and MTB. MAB opsonophagocytic killing activity (OPKA) was examined using SMEG with HL60 and U-937 cells and MTB with U-937 cells. Clearance of MTB from blood was evaluated in Institute of Cancer Research (ICR) mice given opsonic anti-MTB MABs or saline (control) 24 h prior to intravenous infusion with 10⁸ CFUs gamma-irradiated MTB (HN878). MTB levels in murine blood collected 0.25, 4 and 24 h post-challenge were assessed by qPCR. MAB binding to peptidoglycan (PGN) was examined by ELISA using PGN cell wall mixture and ultra-pure PGN. Two MABs (GG9 and JG7) bound to killed and live SMEG and MTB (susceptible and resistant), and promoted OPKA with live MTB. MAB JG7 significantly enhanced OPKA of MTB. Both MABs significantly enhanced clearance of killed MTB from murine blood at 4 and 24 h as measured by qPCR. These opsonic MABs bound to PGN, a major cell wall constituent. Anti-MTB MABs that promote bactericidal phagocytic activity of MTB and enhance clearance of killed MTB from the blood, may offer an immunotherapeutic approach for treatment of MTB bacteremia or sepsis, and augment treatment of multi-drug resistant (MDR) or extensively drug resistant (XDR) TB.
    Keywords Mycobacterium smegmatis ; Mycobacterium tuberculosis ; bacteremia ; blood ; cell walls ; immunotherapy ; intravenous injection ; mice ; models ; multiple drug resistance ; peptidoglycans ; phagocytosis ; tuberculosis ; Biotechnology ; Immunology ; Microbiology ; Molecular biology ; Systems biology ; Monoclonal antibodies ; Opsonophagocytosis ; MAB binding activity ; MTB clearance ; qPCR
    Language English
    Dates of publication 2019-09
    Publishing place Elsevier Ltd
    Document type Article ; Online
    Note NAL-AP-2-clean ; Use and reproduction
    ZDB-ID 2835763-2
    ISSN 2405-8440
    ISSN 2405-8440
    DOI 10.1016/j.heliyon.2019.e02260
    Database NAL-Catalogue (AGRICOLA)

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