Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

New method to diagnose sepsis is faster, cheaper

19.11.2013
A new method could cut hours off the time it takes to diagnose blood infections while also eliminating the need for complicated manual processing and expensive equipment, according to a report to be published in mBio®, the online open-access journal of the American Society for Microbiology, on November 19.

The method combines a selective lysis step in which blood cells in the sample are destroyed, a centrifugation step to collect any bacteria or fungi in the sample, and a fluorescence step that analyzes the particular fingerprint of any pathogens present in the sample.


This is a graphic included with the press release.

Credit: John Walsh et al.

Tests show the method correctly identifies the species of bacteria or fungi in 96.5 % of positive blood culture samples, crucial information for doctors to provide the appropriate drugs for their patients.

"The primary benefit of getting a rapid identification is making sure the patient is on the right [antibiotic] therapy and to quickly make any needed adjustments to the initial therapy," says co-author John Walsh of bioMérieux, Inc. in Durham, North Carolina. Patients with bloodstream infections are usually in very serious condition, says Walsh, and faster identification of the organism causing the infection can help get patients on the most effective antibiotics faster and save lives. Proper diagnosis is also important from the perspective of antibiotic stewardship: using more appropriate, targeted antibiotics reduces the risk of contributing to the spread of resistance to broad-spectrum antibiotics.

Walsh says the current standard approach to diagnosing bloodstream infections, Gram staining and overnight sub-culture followed by phenotypic ID tests, have limitations that can prevent timely treatment. Gram staining provides early, low level information about the type of microorganism present, but it sometimes takes hours to deliver a result, and technicians can make mistakes in the process that provide misleading results. Other more specific identification methods are also available for diagnosis, but they can take at least a day or two to produce results and many require expensive equipment.

In the technique developed by Walsh et al., a sample of positive blood culture is treated with lysis buffer to "pop" the blood cells, then transferred to a specialized optical tube. The tube is centrifuged, which drives bacteria or fungi, which are denser than the solution, down through a liquid density cushion to form a pellet at the bottom of the tube.

Then comes the intrinsic fluorescence spectroscopy (IFS): the microbial pellet is irradiated with light ranging from the deep ultraviolet to infrared, which excites certain organic molecules in the microorganisms, including tryptophan, NADH, FAD, porphyrins, and others, and causes them to fluoresce in a characteristic way depending on the identity of the microbe. The exact pattern of fluorescence is compared with a database of 37 of the most common known pathogens to identify the organism present.

"We're using intrinsic fluorescence to identify the microorganisms. It's an innate property of most living organisms. Because it's intrinsic, no reagents are needed for the identification step," which removes many of the opportunities for mistakes and lowers test costs, says Walsh.

Testing in a controlled laboratory study shows the method can correctly identify the species in 96.5% of all test samples, and in the 2.7% of samples for which no species identity was provided, the system was able to correctly identify 67% to the family level, which is often enough information to select an effective therapy. Among over a thousand samples tested, the method never gave an incorrect result as to the family level or the Gram type.

Walsh says the research and development team in Durham is actively working on automating the system with robotics to make it a fully hands-off process. Blood cultures grow in their own time, often producing a positive result at an inconvenient time of the day for clinical labs, he points out, so automation could speed up diagnosis significantly.

"Our vision is to have a system that will automatically identify the blood culture isolate within 15 minutes of the culture being called positive," says Walsh. If a culture is positive at 2 AM, he says, automating this method could make it possible to identify the organism by 2:15 AM and send an electronic report to a patient's physician. They hope be begin testing and evaluating the feasibility of an automated form of the system in a clinical environment within months.

Rapid ID Procedure: An overall schematic of the simple three-step process (lyse-spin-read) is given in Figures 1a-c. Briefly, a 2.0 mL sample of warm (35-37oC) positive broth is removed from the test blood culture bottle and added to 1.0 mL of warm (35-37oC) selective lysis buffer (0.45% w/v Brij-O10 in 0.3M CAPS, pH 11.7), contained in a 15 mL screw capped polypropylene tube. The mixture was vortexed for 5 seconds at maximum speed and then placed in a 35-37oC waterbath for 60 seconds. After an additional 1-2 second vortex, 1.0 mL of the lysate was removed and layered onto a single 5/16 inch diameter polypropylene ball (CIC Ball Co.) floating on the surface of 0.5 mL of a solution of 24% w/v cesium chloride + 0.005% w/v Pluronic F-108 + 10 mM HEPES, pH 7.4 contained within an optical micro-centrifuge tube. The polypropylene ball was used to control the layering process and create an undisturbed interface. The tube was sealed with a screw-cap and centrifuged for 2 minutes at 10,000 rpm at 20-25oC in a microcentrifuge with A-8-11 swing-out rotor.

mBio® is an open access online journal published by the American Society for Microbiology to make microbiology research broadly accessible. The focus of the journal is on rapid publication of cutting-edge research spanning the entire spectrum of microbiology and related fields. It can be found online at http://mbio.asm.org.

The American Society for Microbiology is the largest single life science society, composed of over 39,000 scientists and health professionals. ASM's mission is to advance the microbiological sciences as a vehicle for understanding life processes and to apply and communicate this knowledge for the improvement of health and environmental and economic well-being worldwide.

Garth Hogan | EurekAlert!
Further information:
http://www.asmusa.org
http://mbio.asm.org

More articles from Life Sciences:

nachricht Flow of cerebrospinal fluid regulates neural stem cell division
22.05.2018 | Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt

nachricht Chemists at FAU successfully demonstrate imine hydrogenation with inexpensive main group metal
22.05.2018 | Friedrich-Alexander-Universität Erlangen-Nürnberg

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

Die letzten 5 Focus-News des innovations-reports im Überblick:

Im Focus: Explanation for puzzling quantum oscillations has been found

So-called quantum many-body scars allow quantum systems to stay out of equilibrium much longer, explaining experiment | Study published in Nature Physics

Recently, researchers from Harvard and MIT succeeded in trapping a record 53 atoms and individually controlling their quantum state, realizing what is called a...

Im Focus: Dozens of binaries from Milky Way's globular clusters could be detectable by LISA

Next-generation gravitational wave detector in space will complement LIGO on Earth

The historic first detection of gravitational waves from colliding black holes far outside our galaxy opened a new window to understanding the universe. A...

Im Focus: Entangled atoms shine in unison

A team led by Austrian experimental physicist Rainer Blatt has succeeded in characterizing the quantum entanglement of two spatially separated atoms by observing their light emission. This fundamental demonstration could lead to the development of highly sensitive optical gradiometers for the precise measurement of the gravitational field or the earth's magnetic field.

The age of quantum technology has long been heralded. Decades of research into the quantum world have led to the development of methods that make it possible...

Im Focus: Computer-Designed Customized Regenerative Heart Valves

Cardiovascular tissue engineering aims to treat heart disease with prostheses that grow and regenerate. Now, researchers from the University of Zurich, the Technical University Eindhoven and the Charité Berlin have successfully implanted regenerative heart valves, designed with the aid of computer simulations, into sheep for the first time.

Producing living tissue or organs based on human cells is one of the main research fields in regenerative medicine. Tissue engineering, which involves growing...

Im Focus: Light-induced superconductivity under high pressure

A team of scientists of the Max Planck Institute for the Structure and Dynamics of Matter (MPSD) at the Center for Free-Electron Laser Science in Hamburg investigated optically-induced superconductivity in the alkali-doped fulleride K3C60under high external pressures. This study allowed, on one hand, to uniquely assess the nature of the transient state as a superconducting phase. In addition, it unveiled the possibility to induce superconductivity in K3C60 at temperatures far above the -170 degrees Celsius hypothesized previously, and rather all the way to room temperature. The paper by Cantaluppi et al has been published in Nature Physics.

Unlike ordinary metals, superconductors have the unique capability of transporting electrical currents without any loss. Nowadays, their technological...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

VideoLinks
Industry & Economy
Event News

Save the date: Forum European Neuroscience – 07-11 July 2018 in Berlin, Germany

02.05.2018 | Event News

Invitation to the upcoming "Current Topics in Bioinformatics: Big Data in Genomics and Medicine"

13.04.2018 | Event News

Unique scope of UV LED technologies and applications presented in Berlin: ICULTA-2018

12.04.2018 | Event News

 
Latest News

Supersonic waves may help electronics beat the heat

18.05.2018 | Power and Electrical Engineering

Keeping a Close Eye on Ice Loss

18.05.2018 | Information Technology

CrowdWater: An App for Flood Research

18.05.2018 | Information Technology

VideoLinks
Science & Research
Overview of more VideoLinks >>>