Academic Journal

Multilabel hybridization probes for sequence-specific detection of sepsis-related drug resistance genes in plasmids

التفاصيل البيبلوغرافية
العنوان: Multilabel hybridization probes for sequence-specific detection of sepsis-related drug resistance genes in plasmids
المؤلفون: Robert L. Hanson, Elaine Lazalde, Radim Knob, David H. Harris, Yesman Akuoko, Jacob B. Nielsen, Adam T. Woolley
المصدر: Talanta Open, Vol 3, Iss , Pp 100034- (2021)
بيانات النشر: Elsevier, 2021.
سنة النشر: 2021
المجموعة: LCC:Analytical chemistry
مصطلحات موضوعية: DNA hybridization, Sequence-specific hybridization, Multilabel fluorescence, Drug resistance, Plasmid detection, Analytical chemistry, QD71-142
الوصف: Emerging antimicrobial drug resistance is increasing the complexity involved in treating critical conditions such as bacterial induced sepsis. Methods for diagnosing specific drug resistance tend to be rapid or sensitive, but not both. Detection methods like sequence-specific single-molecule analysis could address this concern if they could be adapted to work on smaller targets similar to those produced in traditional clinical situations. In this work we demonstrate that a 120 bp double stranded polynucleotide with an overhanging single stranded 25 bp probe sequence can be created by immobilizing DNA with a biotin/streptavidin magnetic bead system, labeling with SYBR Gold, and rinsing the excess away while the probe retains multiple fluorophores. These probes with multiple fluorophores can then be used to label a bacterial plasmid target in a sequence-specific manner. These probes enabled the detection of 1 pM plasmid samples containing a portion of an antibiotic resistance gene sequence. This system shows the possibility of improving capture and fluorescence labeling of small nucleic acid fragments, generating lower limits of detection for clinically relevant samples while maintaining rapid processing times.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 2666-8319
Relation: http://www.sciencedirect.com/science/article/pii/S2666831921000047; https://doaj.org/toc/2666-8319
DOI: 10.1016/j.talo.2021.100034
URL الوصول: https://doaj.org/article/f8854e44d012440aa279f60cc27bbca5
رقم الانضمام: edsdoj.f8854e44d012440aa279f60cc27bbca5
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:26668319
DOI:10.1016/j.talo.2021.100034