Scientists of the German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ) and the Universities of Heidelberg and Bielefeld, Germany, have developed a highly sensitive test for detecting this genetic alteration at the level of a single molecule, thus providing information about the resistance status of an infected person.
Many resistances to antibiotics are based on specific mutations in the genetic material of the infectious agents. In the case of life-threatening infections it is vital to determine rapidly which medication will work for the patient. However, commonly used methods of resistance detection are too time-consuming, particularly with microorganisms such as tuberculosis bacteria, which grow very slowly in the culture dish.
Scientists headed by Dr. Jens-Peter Knemeyer of the Division of Functional Genome Analysis at the DKFZ have combined a hybridization method, where small DNA probes bind highly specifically and exclusively to the mutated gene sequence, with confocal microscopy technology. The DNA probes are coupled to a fluorescent dye that flashes under laser light. However, this light signal is emitted only if the probe attaches to the target sequence in the bacterial genetic material. ‘Unbound’ probe molecules do not emit a signal. Each of these tiny light flashes that occur when the probe and the target molecule bind to each other, detects a single mutated DNA molecule.
By measuring the duration and decay times of the light flashes, the researchers distinguish between real measurement results and the ubiquitous background fluorescence: Due to chemical properties of the molecules involved, spontaneous fluorescence decays much more quickly than the signal emitted by the dye-labeled probe.
Detection of resistance causing point mutations in the genetic material of the tuberculosis bacterium is just one of numerous possible applications of the new method called single-molecule fluorescence spectroscopy. The method has a big advantage: Instead of recording light flashes in a sample solution, as is done in antibiotic resistance detection, the investigation method can also be used in living cells. Dr. Jörg Hoheisel, head of the Division of Functional Genome Analysis at the DKFZ, explains: “Just as we can detect DNA mutations, we can also use suitable probes to detect all molecules in a cell that are characteristic of a specific disease. Since the test identifies single molecules, it is highly sensitive – but reliable at the same time, because we have an internal control using the decay times.”
Julia Rautenstrauch | alfa
Rainbow colors reveal cell history: Uncovering β-cell heterogeneity
22.09.2017 | DFG-Forschungszentrum für Regenerative Therapien TU Dresden
The pyrenoid is a carbon-fixing liquid droplet
22.09.2017 | Max-Planck-Institut für Biochemie
Plants and algae use the enzyme Rubisco to fix carbon dioxide, removing it from the atmosphere and converting it into biomass. Algae have figured out a way to increase the efficiency of carbon fixation. They gather most of their Rubisco into a ball-shaped microcompartment called the pyrenoid, which they flood with a high local concentration of carbon dioxide. A team of scientists at Princeton University, the Carnegie Institution for Science, Stanford University and the Max Plank Institute of Biochemistry have unravelled the mysteries of how the pyrenoid is assembled. These insights can help to engineer crops that remove more carbon dioxide from the atmosphere while producing more food.
A warming planet
Our brains house extremely complex neuronal circuits, whose detailed structures are still largely unknown. This is especially true for the so-called cerebral cortex of mammals, where among other things vision, thoughts or spatial orientation are being computed. Here the rules by which nerve cells are connected to each other are only partly understood. A team of scientists around Moritz Helmstaedter at the Frankfiurt Max Planck Institute for Brain Research and Helene Schmidt (Humboldt University in Berlin) have now discovered a surprisingly precise nerve cell connectivity pattern in the part of the cerebral cortex that is responsible for orienting the individual animal or human in space.
The researchers report online in Nature (Schmidt et al., 2017. Axonal synapse sorting in medial entorhinal cortex, DOI: 10.1038/nature24005) that synapses in...
Whispering gallery mode (WGM) resonators are used to make tiny micro-lasers, sensors, switches, routers and other devices. These tiny structures rely on a...
Using ultrafast flashes of laser and x-ray radiation, scientists at the Max Planck Institute of Quantum Optics (Garching, Germany) took snapshots of the briefest electron motion inside a solid material to date. The electron motion lasted only 750 billionths of the billionth of a second before it fainted, setting a new record of human capability to capture ultrafast processes inside solids!
When x-rays shine onto solid materials or large molecules, an electron is pushed away from its original place near the nucleus of the atom, leaving a hole...
For the first time, physicists have successfully imaged spiral magnetic ordering in a multiferroic material. These materials are considered highly promising candidates for future data storage media. The researchers were able to prove their findings using unique quantum sensors that were developed at Basel University and that can analyze electromagnetic fields on the nanometer scale. The results – obtained by scientists from the University of Basel’s Department of Physics, the Swiss Nanoscience Institute, the University of Montpellier and several laboratories from University Paris-Saclay – were recently published in the journal Nature.
Multiferroics are materials that simultaneously react to electric and magnetic fields. These two properties are rarely found together, and their combined...
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