A5105873481- Bacteriophages and microbial interactions
- Bacterial Identification and Susceptibility Testing
- Advanced biosensing and bioanalysis techniques
- Antimicrobial Resistance in Staphylococcus
- DNA and Nucleic Acid Chemistry
- Infective Endocarditis Diagnosis and Management
- Bacterial biofilms and quorum sensing
- Vibrio bacteria research studies
- Stress Responses and Cortisol
- Antibiotic Resistance in Bacteria
- Mycobacterium research and diagnosis
- Water Treatment and Disinfection
- Fungal Infections and Studies
- Molecular Biology Techniques and Applications
- Virus-based gene therapy research
- Microbial Fuel Cells and Bioremediation
- Antifungal resistance and susceptibility
- Growth Hormone and Insulin-like Growth Factors
- Endoplasmic Reticulum Stress and Disease
- Blood groups and transfusion
- Hormonal Regulation and Hypertension
- Diphtheria, Corynebacterium, and Tetanus
- CRISPR and Genetic Engineering
- Neonatal and Maternal Infections
- Wastewater Treatment and Nitrogen Removal
Boston University
2002
Boston Children's Hospital
1992
Harvard University
1990
Brigham and Women's Hospital
1990
Molecular beacons are sensitive fluorescent probes hybridizing selectively to designated DNA and RNA targets. They have recently become practical tools for quantitative real-time monitoring of single-stranded nucleic acids. Here, we comparatively study the performance a variety such probes, stemless stem-containing PNA (peptide acid) beacons, in Tris-buffer solutions containing various concentrations NaCl MgCl2. We demonstrate that different molecular respond differently change salt...
We evaluated the performance of Candida albicans/Candida glabrata peptide nucleic acid fluorescent in situ hybridization (PNA FISH) method, a rapid two-color assay for detection C. albicans and glabrata, multicenter study. The is designed use directly from positive blood culture bottles FISH format. Intact, fixed cells are labeled green (C. albicans) or red glabrata) by rRNA fluorophore-labeled PNA probes. Results available <3 h after cultures signal positive. An evaluation 197 routine newly...
A novel rapid peptide nucleic acid fluorescence in situ hybridization (FISH) method, Staphylococcus QuickFISH, for the direct detection of species from positive blood culture bottles was evaluated a multicenter clinical study. The method utilizes microscope slide with predeposited positive- and negative-control organisms self-reporting 15-min step, which eliminates need wash step. Five laboratories tested 722 containing gram-positive cocci clusters. sensitivities aureus coagulase-negative...
We report here on the hybridization of peptide nucleic acid (PNA)-based molecular beacons (MB) directly to duplex DNA sites locally exposed by PNA openers. Two stemless were tested, both featuring same recognition sequence and fluorophore-quencher pair (Fluorescein DABCYL, respectively) but differing in arrangement these groups net electrostatic charge. It was found that one beacon rapidly hybridized, with aid openers, its complementary target within at ambient conditions via formation a...
The utility of peptide nucleic acid fluorescence in situ hybridization (PNA FISH) for the detection Acinetobacter spp. and Pseudomonas aeruginosa was evaluated on broth suspensions spiked blood cultures ATCC strains clinical isolates with select gram-negative rods. After testing 60 isolates, PNA FISH had a sensitivity specificity 100% 100%, respectively, 95%, P. aeruginosa. able to detect both pathogens simultaneously directly from cultures.
We report a new fluorogenic method for sealed-tube PCR analysis using quencher-labeled peptide nucleic acid (Q-PNA) probe. The Q-PNA hybridizes to complementary tag sequence located at the 5' end of fluorophore-labeled oligonucleotide primer, quenching primer's fluorescence. Incorporation primer into doublestranded amplicon causes displacement such that fluorescence sample is direct indication concentration. able quench multiple primers bearing distinct fluorophores in single reaction. show...
This study evaluated a novel peptide nucleic acid (PNA) probe targeting region of the 23S rRNA gene Klebsiella pneumoniae by fluorescence in situ hybridization (FISH). Analytical performance was determined using 39 reference strains and other well-characterized spp. Enterobacter aerogenes . The found to be specific for K. complex ( including ozaenae variicola ). diagnostic accuracy with 264 blood cultures containing Gram-negative rods. Using conventional identification as reference,...
We have studied the effect of protein kinase-C activation on regulation CRH gene expression in human hepatoma cell line NPLC/PRF/5 (NPLC), only known to express endogenous gene. Incubation NPLC cells with 100 nM 12-O-tetradecanoyl phorbol 13-acetate (TPA), a ester that activates kinase-C, resulted rapid (1-h) and prolonged (72-h) increase mRNA levels, maximum 16-fold observed at 24 h. In addition, TPA treatment increased size by approximately nucleotides. This increase, which was blocked...
The applicability of the PNA FISH (peptide nucleic acid fluorescence in situ hybridization) method for detection Streptococcus agalactiae [group B streptococci (GBS)] from swab samples was evaluated. Three swab-sample-processing protocols with different time-to-result (TTR) values were compared: (i) direct smearing fresh swabs onto microscope slides (n=153, TTR 2.5 h), (ii) further extraction and concentration cells these same 2.7 (iii) short-term LIM broth enrichment culture incubation (7...
The objective of this study was to develop a novel peptide nucleic acid (PNA) probe for Stenotrophomonas maltophilia identification by fluorescence in situ hybridization (FISH). evaluated using 33 human and veterinary clinical S. isolates 45 reference strains representing common bacterial species the respiratory tract. displayed 100% sensitivity specificity on pure cultures allowed detection sputum from cystic fibrosis patients. limit 10(4) CFU/mL spiked tracheal aspirate bronchoalveolar...
Triplex gene editing relies on binding a stable peptide nucleic acid (PNA) sequence to chromosomal target, which alters the helical structure of DNA stimulate site-specific recombination with single-strand (ssDNA) donor template and elicits correction. Here, we assessed whether codelivery PNA encapsulated in Poly Lactic-co-Glycolic Acid (PLGA)-based nanoparticles can correct sickle cell disease x-linked severe combined immunodeficiency. However, through this process have identified...