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  1. Article: Rapid and specific detection of Mycobacterium tuberculosis from acid-fast bacillus smear-positive respiratory specimens and BacT/ALERT MP culture bottles by using fluorogenic probes and real-time PCR.

    Miller, Nancimae / Cleary, Tim / Kraus, Günter / Young, Andrea K / Spruill, Gina / Hnatyszyn, H James

    Journal of clinical microbiology

    2001  Volume 40, Issue 11, Page(s) 4143–4147

    Abstract: A real-time PCR assay using the LightCycler (LC) instrument for the specific identification of Mycobacterium tuberculosis complex (MTB) was employed to detect organisms in 135 acid-fast bacillus (AFB) smear-positive respiratory specimens and in 232 BacT/ ... ...

    Abstract A real-time PCR assay using the LightCycler (LC) instrument for the specific identification of Mycobacterium tuberculosis complex (MTB) was employed to detect organisms in 135 acid-fast bacillus (AFB) smear-positive respiratory specimens and in 232 BacT/ALERT MP (MP) culture bottles of respiratory specimens. The LC PCR assay was directed at the amplification of the internal transcribed spacer region of the Mycobacterium genome with real-time detection using fluorescence resonance energy transfer probes specific for MTB. The results from the respiratory specimens were compared to those from the Amplicor M. tuberculosis PCR test. Specimens from MP culture bottles were analyzed by Accuprobe and conventional identification methods. MTB was cultured from 105 (77.7%) respiratory AFB smear-positive specimens; 103 of these samples were positive by LC PCR and Amplicor PCR. Two samples negative in the LC assay contained rare numbers of organisms; both were positive in the Amplicor assay. Two separate samples negative by Amplicor PCR contained low and moderate numbers of AFB, respectively, and both of these were positive in the LC assay. There were 30 AFB smear-positive respiratory specimens that grew mycobacteria other than tuberculosis (MOTT), and all tested negative in both assays. Of the 231 MP culture bottles, 114 cultures were positive for MTB and all were positive by the LC assay. The remaining 117 culture bottles were negative in the LC assay and grew various MOTT. This real-time MTB assay is sensitive and specific; a result was available within 1 h of having a DNA sample available for testing.
    MeSH term(s) Bacterial Typing Techniques ; Culture Media ; Fluorescence Resonance Energy Transfer ; Fluorescent Dyes ; Humans ; Mycobacterium tuberculosis/genetics ; Mycobacterium tuberculosis/growth & development ; Mycobacterium tuberculosis/isolation & purification ; Polymerase Chain Reaction/methods ; Reagent Kits, Diagnostic ; Respiratory System/microbiology ; Sensitivity and Specificity ; Specimen Handling/methods ; Temperature ; Time Factors ; Tuberculosis, Pulmonary/microbiology
    Chemical Substances Culture Media ; Fluorescent Dyes ; Reagent Kits, Diagnostic
    Language English
    Publishing date 2001-10-13
    Publishing country United States
    Document type Evaluation Study ; Journal Article ; 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.40.11.4143-4147.2002
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1188: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

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  2. Article: Cross-clade inhibition of HIV-1 replication and cytopathology by using RNase P-associated external guide sequences.

    Kraus, Gunter / Geffin, Rebeca / Spruill, Gina / Young, Andrea K / Seivright, Rachel / Cardona, Diana / Burzawa, Jennifer / Hnatyszyn, H James

    Proceedings of the National Academy of Sciences of the United States of America

    2002  Volume 99, Issue 6, Page(s) 3406–3411

    Abstract: RNase P complexes have been proposed as a novel RNA-based gene interference strategy to inhibit gene expression in human malignancies and infectious diseases. This approach is based on the sequence-specific design of an external guide sequence (EGS) RNA ... ...

    Abstract RNase P complexes have been proposed as a novel RNA-based gene interference strategy to inhibit gene expression in human malignancies and infectious diseases. This approach is based on the sequence-specific design of an external guide sequence (EGS) RNA molecule that can specifically hybridize to almost any complementary target mRNA and facilitate its cleavage by the RNase P enzyme component. We designed a truncated RNase P-associated EGS molecule to specifically recognize the U5 region of HIV-1 mRNA and mediate cleavage of hybridized mRNA by the RNase P enzyme. Genes encoding for this U5-EGS (560) molecule, as well as a U5 EGS (560D) antisense control, were cloned into retroviral plasmids and transferred into a CD4(+) T cell line. Transfected cells were exposed to increasing concentrations of HIV-1 clinical isolates from clades A, B, C, and F. Heterogeneous cultures of CD4(+) T cells expressing the U5 EGS (560) molecule were observed to maintain CD4 levels, were devoid of cytopathology, and did not produce HIV p24 gag antigen through 30 days after exposure to all HIV-1 clades at a multiplicity of infection of 0.01. Identical cells expressing the U5 EGS (560D) antisense control molecule underwent a loss of CD4 expression, produced elevated levels of HIV-1, and formed large syncytia similar to untreated cells. When the viral inoculum was increased at the time of exposure (multiplicity of infection = 0.05), the inhibitory effect of the U5 EGS (560) molecule was overwhelmed, but viral-mediated cytopathology and particle production were delayed compared with control cell populations. Viral replication and cytopathology associated with infection of multiple HIV-1 clades can be effectively inhibited in CD4(+) cells expressing the RNase P-associated U5 EGS (560) molecule.
    MeSH term(s) CD4-Positive T-Lymphocytes/metabolism ; CD4-Positive T-Lymphocytes/virology ; Cell Line ; Cytopathogenic Effect, Viral ; DNA, Complementary/genetics ; DNA, Viral/biosynthesis ; DNA, Viral/genetics ; Endoribonucleases/biosynthesis ; Endoribonucleases/genetics ; Endoribonucleases/metabolism ; Flow Cytometry ; HIV Core Protein p24/biosynthesis ; HIV Core Protein p24/genetics ; HIV-1/classification ; HIV-1/genetics ; HIV-1/metabolism ; HIV-1/physiology ; Humans ; Plasmids/genetics ; Proviruses/genetics ; RNA, Catalytic/biosynthesis ; RNA, Catalytic/genetics ; RNA, Catalytic/metabolism ; Retroviridae/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Ribonuclease P ; Substrate Specificity ; Transfection ; Virus Replication ; RNA, Small Untranslated
    Chemical Substances DNA, Complementary ; DNA, Viral ; HIV Core Protein p24 ; RNA, Catalytic ; Endoribonucleases (EC 3.1.-) ; RPP14 protein, human (EC 3.1.26.5) ; Ribonuclease P (EC 3.1.26.5) ; RNA, Small Untranslated
    Language English
    Publishing date 2002-06-05
    Publishing country United States
    Document type Journal Article
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.052651199
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1148: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

    Order via subito

    This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

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