สืบค้นงานวิจัย
Rapid molecular detection of multidrug- resistant tuberculosis by PCR-nucleic acid lateral flow immunoassay
Kamphee H. - ไม่ระบุหน่วยงาน
ชื่อเรื่อง (EN): Rapid molecular detection of multidrug- resistant tuberculosis by PCR-nucleic acid lateral flow immunoassay
ผู้แต่ง / หัวหน้าโครงการ (EN): Kamphee H.
บทคัดย่อ (EN): Several existing molecular tests for multidrug-resistant tuberculosis (MDR-TB) are limited by complexity and cost, hindering their widespread application. The objective of this proof of concept study was to develop a simple Nucleic Acid Lateral Flow (NALF) immunoassay as a potential diagnostic alternative, to complement conventional PCR, for the rapid molecular detection of MDR-TB. The NALF device was designed using antibodies for the indirect detection of labeled PCR amplification products. Multiplex PCR was optimized to permit the simultaneous detection of the drug resistant determining mutations in the 81-bp hot spot region of the rpoB gene (rifampicin resistance), while semi-nested PCR was optimized for the S315T mutation detection in the katG gene (isoniazid resistance). The amplification process additionally targeted a conserved region of the genes as Mycobacterium tuberculosis (Mtb) DNA control. The optimized conditions were validated with the H37Rv wild-type (WT) Mtb isolate and Mtb isolates with known mutations (MT) within the rpoB and katG genes. Results indicate the correct identification of WT (drug susceptible) and MT (drug resistant) Mtb isolates, with the least limit of detection (LOD) being 10 4 genomic copies per PCR reaction. NALF is a simple, rapid and low-cost device suitable for low resource settings where conventional PCR is already employed on a regular basis. Moreover, the use of antibodybased NALF to target primer-labels, without the requirement for DNA hybridization, renders the device generic, which could easily be adapted for the molecular diagnosis of other infectious and non-infectious diseases requiring nucleic acid detection. Copyright: © 2015 Kamphee et al.
บทคัดย่อ: ไม่พบข้อมูลจากหน่วยงานต้นทาง
ภาษา (EN): en
เอกสารแนบ (EN): https://www.scopus.com/inward/record.uri?eid=2-s2.0-84944754738&doi=10.1371%2fjournal.pone.0137791&partnerID=40&md5=9bbdabf90a65c3213429e964fdb95e42
เผยแพร่โดย (EN): มหาวิทยาลัยมหิดล
คำสำคัญ (EN): Tuberculosis, Multidrug-Resistant
เจ้าของลิขสิทธิ์ (EN): มหาวิทยาลัยมหิดล
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Rapid molecular detection of multidrug- resistant tuberculosis by PCR-nucleic acid lateral flow immunoassay
Kamphee H.
มหาวิทยาลัยมหิดล
ไม่ระบุวันที่เผยแพร่
Rapid detection of tuberculosis resistant to isoniazid and rifampicin by real-time PCR and melting curve analysis Detection of Mycobacterium tuberculosis and Its Rifampicin Resistance by Automated Nucleic Acid Amplification Test Development of competitive lateral flow immunoassay coupled with silver enhancement for simple and sensitive salivary cortisol detection Antibody-conjugated ferromagnetic nanoparticles with lateral flow test strip assay for rapid detection of Campylobacter jejuni in poultry samples Rapid Molecular Identification and Detection of Corynespora cassiicola of Hevea brasiliensis Risk factors for multidrug-resistant tuberculosis among patients with pulmonary tuberculosis at the central chest institute of Thailand Prevalence and extent of heteroresistance by next generation sequencing of multidrug-resistant tuberculosis Development of molecular approach based on PCR assay for detection of histamine producing bacteria A silicon nitride ISFET based immunosensor for Ag85B detection of tuberculosis Genotypic diversity of multidrug-, quinolone- and extensively drug-resistant Mycobacterium tuberculosis isolates in Thailand
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