A straightforward DOPE (double labeling of oligonucleotide probes)-FISH (fluorescence in situ hybridization) method for simultaneous multicolor detection of six microbial populations.

Applied and Environmental Microbiology
Faris BehnamK Stoecker

Abstract

Fluorescence in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes is an essential tool for the cultivation-independent identification of microbes within environmental and clinical samples. However, one of the major constraints of conventional FISH is the very limited number of different target organisms that can be detected simultaneously with standard epifluorescence or confocal laser scanning microscopy. Recently, this limitation has been overcome via an elegant approach termed combinatorial labeling and spectral imaging FISH (CLASI-FISH) (23). This technique, however, suffers compared to conventional FISH from an inherent loss in sensitivity and potential probe binding biases caused by the competition of two differentially labeled oligonucleotide probes for the same target site. Here we demonstrate that the application of multicolored, double-labeled oligonucleotide probes enables the simultaneous detection of up to six microbial target populations in a straightforward and robust manner with higher sensitivity and less bias. Thus, this newly developed technique should be an attractive option for all researchers interested in applying conventional FISH methods for the study of microbial communities.

References

Jun 1, 1996·Journal of Bacteriology·R AmannK H Schleifer
Oct 6, 1997·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·E J SpeelA H Hopman
Jun 1, 2002·Applied and Environmental Microbiology·Annelie PernthalerRudolf Amann
Jul 2, 2003·Current Opinion in Microbiology·Michael WagnerHolger Daims
Nov 25, 2003·Analytical Chemistry·Thomas L FareYanqun Wang
Dec 3, 2004·Applied and Environmental Microbiology·L Safak Yilmaz, Daniel R Noguera
Jan 5, 2006·Applied and Environmental Microbiology·L Safak YilmazDaniel R Noguera
Jan 21, 2006·Environmental Microbiology·Holger DaimsMichael Wagner
Nov 14, 2006·Nucleic Acids Research·Alexander LoyMatthias Horn
Mar 3, 2007·Methods in Molecular Biology·Annelie Pernthaler, Jakob Pernthaler
Jun 8, 2007·Microbiology and Molecular Biology Reviews : MMBR·Michael W TaylorMichael Wagner
Nov 29, 2007·Diagnostic Molecular Pathology : the American Journal of Surgical Pathology, Part B·Raymond R TubbsJoseph P Crowe
Apr 17, 2008·Nature Reviews. Microbiology·Rudolf Amann, Bernhard M Fuchs
Feb 18, 2011·Proceedings of the National Academy of Sciences of the United States of America·Alex M ValmGary G Borisy
Mar 30, 2011·Environmental Microbiology·Nicole S Webster, Michael W Taylor
Aug 1, 2010·Environmental Microbiology·Nicole S WebsterMichael Wagner
Nov 15, 2011·Current Opinion in Biotechnology·Michael Wagner, Susanne Haider
Dec 20, 2011·Environmental Microbiology·Regina RadaxChrista Schleper

❮ Previous
Next ❯

Citations

Jul 10, 2013·Environmental Science and Pollution Research International·Kerstin Toebe
Jul 31, 2013·Applied and Environmental Microbiology·Robert AlmstrandMalte Hermansson
May 8, 2015·Folia Microbiologica·Monique KerstensPaul Cos
Oct 18, 2015·Applied and Environmental Microbiology·Mario P SchimakBernhard M Fuchs
Aug 18, 2018·Environmental Microbiology·Pedro J Cabello-YevesFrancisco Rodriguez-Valera
Sep 17, 2020·Expert Opinion on Biological Therapy·Varun S Wakade, Pravin Shende
Jul 7, 2017·Frontiers in Microbiology·Pedro J Cabello-YevesFrancisco Rodriguez-Valera
Mar 6, 2018·Journal of Oral Microbiology·Ingar Olsen
Sep 10, 2015·Microbiology Spectrum·Michael J FranklinBrian Bothner
Jun 22, 2021·Current Opinion in Biomedical Engineering·Hao ShiIwijn De Vlaminck
May 15, 2018·ACS Sensors·Mandana AmiriSabine Szunerits

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.