Abstract

Background: Tuberculosis accounted for 8.7 million new cases in 2011 and continues to be one of the leading human infectious diseases. Burdensome is the increasing rate of multi-drug resistant tuberculosis (MDRTB) and the difficulties created for treatment and public health control programs, especially in developing countries. Resistance to rifampicin (RIF), a first line antibiotic, is commonly associated with point mutations within the rpoB gene of Mycobacterium tuberculosis (Mtb) whose detection is considered the best early molecular predictor for MDRTB. Gold nanoparticles functionalized with thiol-modified oligonucleotides (Au-nanoprobes) have shown the potential to provide a rapid and sensitive detection method for Mtb and single base alterations associated with antibiotic resistance, namely in rpoB gene associated to RIF resistance.Results: We developed a strategy based on the isothermal amplification of sample DNA (LAMP) coupled to specific Au-nanoprobes capable of identifying members of the Mtb complex (MTBC) and discriminating specific mutations within the rpoB gene. Integration of LAMP and Au-nanoprobe assay allowed to detect MTBC member and identify mutations linked to RIF resistance. A total of 12 biological samples were tested and a 100% specificity and sensitivity was attained.Conclusions: There is an increasing demand for simple, fast and cheap methods for the molecular identification of Mtb and for the detection of molecular tags associated to drug resistance suitable for use at point-of-need. Here we describe such a method, that as the potential to get molecular diagnostic of tuberculosis to remote environments.
Original languageUnknown
Pages (from-to)38
JournalJournal of Nanobiotechnology
Volume11
Issue numberNA
DOIs
Publication statusPublished - 1 Jan 2013

Keywords

    Cite this

    @article{c1384fed479446338b01bbe0c27aedb0,
    title = "Isothermal DNA amplification coupled to Aunanoprobes for detection of mutations associated to Rifampicin resistance in Mycobacterium tuberculosis",
    abstract = "Background: Tuberculosis accounted for 8.7 million new cases in 2011 and continues to be one of the leading human infectious diseases. Burdensome is the increasing rate of multi-drug resistant tuberculosis (MDRTB) and the difficulties created for treatment and public health control programs, especially in developing countries. Resistance to rifampicin (RIF), a first line antibiotic, is commonly associated with point mutations within the rpoB gene of Mycobacterium tuberculosis (Mtb) whose detection is considered the best early molecular predictor for MDRTB. Gold nanoparticles functionalized with thiol-modified oligonucleotides (Au-nanoprobes) have shown the potential to provide a rapid and sensitive detection method for Mtb and single base alterations associated with antibiotic resistance, namely in rpoB gene associated to RIF resistance.Results: We developed a strategy based on the isothermal amplification of sample DNA (LAMP) coupled to specific Au-nanoprobes capable of identifying members of the Mtb complex (MTBC) and discriminating specific mutations within the rpoB gene. Integration of LAMP and Au-nanoprobe assay allowed to detect MTBC member and identify mutations linked to RIF resistance. A total of 12 biological samples were tested and a 100{\%} specificity and sensitivity was attained.Conclusions: There is an increasing demand for simple, fast and cheap methods for the molecular identification of Mtb and for the detection of molecular tags associated to drug resistance suitable for use at point-of-need. Here we describe such a method, that as the potential to get molecular diagnostic of tuberculosis to remote environments.",
    keywords = "PCR, Tuberculosis, MDRTB, Gold nanoparticles, Nanodiagnostics, Rifampicin, DNA isothermal amplification, LAMP",
    author = "Baptista, {Pedro Miguel Ribeiro Viana} and Bettencourt, {Miguel Viveiros} and Couto, {Isabel Maria dos Santos Leit{\~a}o}",
    note = "Sem PDF",
    year = "2013",
    month = "1",
    day = "1",
    doi = "10.1186/1477-3155-11-38",
    language = "Unknown",
    volume = "11",
    pages = "38",
    journal = "Journal of Nanobiotechnology",
    issn = "1477-3155",
    publisher = "BIOMED CENTRAL LTD",
    number = "NA",

    }

    TY - JOUR

    T1 - Isothermal DNA amplification coupled to Aunanoprobes for detection of mutations associated to Rifampicin resistance in Mycobacterium tuberculosis

    AU - Baptista, Pedro Miguel Ribeiro Viana

    AU - Bettencourt, Miguel Viveiros

    AU - Couto, Isabel Maria dos Santos Leitão

    N1 - Sem PDF

    PY - 2013/1/1

    Y1 - 2013/1/1

    N2 - Background: Tuberculosis accounted for 8.7 million new cases in 2011 and continues to be one of the leading human infectious diseases. Burdensome is the increasing rate of multi-drug resistant tuberculosis (MDRTB) and the difficulties created for treatment and public health control programs, especially in developing countries. Resistance to rifampicin (RIF), a first line antibiotic, is commonly associated with point mutations within the rpoB gene of Mycobacterium tuberculosis (Mtb) whose detection is considered the best early molecular predictor for MDRTB. Gold nanoparticles functionalized with thiol-modified oligonucleotides (Au-nanoprobes) have shown the potential to provide a rapid and sensitive detection method for Mtb and single base alterations associated with antibiotic resistance, namely in rpoB gene associated to RIF resistance.Results: We developed a strategy based on the isothermal amplification of sample DNA (LAMP) coupled to specific Au-nanoprobes capable of identifying members of the Mtb complex (MTBC) and discriminating specific mutations within the rpoB gene. Integration of LAMP and Au-nanoprobe assay allowed to detect MTBC member and identify mutations linked to RIF resistance. A total of 12 biological samples were tested and a 100% specificity and sensitivity was attained.Conclusions: There is an increasing demand for simple, fast and cheap methods for the molecular identification of Mtb and for the detection of molecular tags associated to drug resistance suitable for use at point-of-need. Here we describe such a method, that as the potential to get molecular diagnostic of tuberculosis to remote environments.

    AB - Background: Tuberculosis accounted for 8.7 million new cases in 2011 and continues to be one of the leading human infectious diseases. Burdensome is the increasing rate of multi-drug resistant tuberculosis (MDRTB) and the difficulties created for treatment and public health control programs, especially in developing countries. Resistance to rifampicin (RIF), a first line antibiotic, is commonly associated with point mutations within the rpoB gene of Mycobacterium tuberculosis (Mtb) whose detection is considered the best early molecular predictor for MDRTB. Gold nanoparticles functionalized with thiol-modified oligonucleotides (Au-nanoprobes) have shown the potential to provide a rapid and sensitive detection method for Mtb and single base alterations associated with antibiotic resistance, namely in rpoB gene associated to RIF resistance.Results: We developed a strategy based on the isothermal amplification of sample DNA (LAMP) coupled to specific Au-nanoprobes capable of identifying members of the Mtb complex (MTBC) and discriminating specific mutations within the rpoB gene. Integration of LAMP and Au-nanoprobe assay allowed to detect MTBC member and identify mutations linked to RIF resistance. A total of 12 biological samples were tested and a 100% specificity and sensitivity was attained.Conclusions: There is an increasing demand for simple, fast and cheap methods for the molecular identification of Mtb and for the detection of molecular tags associated to drug resistance suitable for use at point-of-need. Here we describe such a method, that as the potential to get molecular diagnostic of tuberculosis to remote environments.

    KW - PCR

    KW - Tuberculosis

    KW - MDRTB

    KW - Gold nanoparticles

    KW - Nanodiagnostics

    KW - Rifampicin

    KW - DNA isothermal amplification

    KW - LAMP

    U2 - 10.1186/1477-3155-11-38

    DO - 10.1186/1477-3155-11-38

    M3 - Article

    VL - 11

    SP - 38

    JO - Journal of Nanobiotechnology

    JF - Journal of Nanobiotechnology

    SN - 1477-3155

    IS - NA

    ER -