Thursday, June 21, 2012

Call for Papers PDA Europe Micro Meeting Feb. 2013

As co-chairs of the 2013 PDA Europe Microbiology Conference, it is our pleasure to invite interested parties to submit abstracts for paper or poster presentations to this annual meeting, exposition and training program. Potential topics to be covered include rapid micro methods, environmental monitoring, biofilms and water systems, contamination control, aseptic processing and regulatory expectations, to name a few.

The conference and exhibition will be held on 26-27 February 2013 in Berlin, Germany. Following the conference, a number of microbiology training programs, including the very popular 2-day intensive rapid micro methods course, will be held on 28 February and 1 March.

The Call for Papers deadline is June 29, so act now to submit your abstract! The link to the submission form is

Thank you for your consideration!

Michael J. Miller and Jette Christensen, Co-chairs, 2013 PDA Europe Microbiology Conference Program Committee

Biochip May Track Flu in Real Time

Researchers at Brown University have created a reliable and fast flu-detection test that can be carried in a first-aid kit and may lead to real-time tracking of influenza.

The prototype device, which isolates influenza RNA using a combination of magnetics and microfluidics and then amplifies and detects probes bound to the RNA, is described in the Journal of Molecular Diagnostics.

In April 2009, the world took notice as reports surfaced of a virus in Mexico that had mutated from pigs and was being passed from human to human. The H1N1 “swine flu,” as the virus was named, circulated worldwide, killing more than 18,000 people, according to the World Health Organization. The Centers for Disease Control and Prevention in the U.S. said it was the first global pandemic in more than four decades.

Swine flu won’t be the last viral mutation to cause a worldwide stir. One way to contain the next outbreak is by administering tests at the infection’s source and pinpointing and tracking the pathogen’s spread in real time. But such efforts have been stymied by devices that are costly, unwieldy, and unreliable.

Now, biomedical engineers have developed a biochip that can detect the presence of influenza by zeroing in on the specific RNA sequence and then using tiny magnets in a tube to separate the flu-ridden sequence from the rest of the RNA strand.

The result: A reliable, fast prototype of a flu-detection test that potentially can be carried in a first-aid kit and used as easily as an iPhone.

“We wanted to make something simple,” says Anubhav Tripathi, associate professor of engineering at Brown University and the paper’s corresponding author. “It’s a low-cost device for active, on-site detection, whether it’s influenza, HIV, or TB (tuberculosis).”

The Brown assay is called SMART, which stands for “A Simple Method for Amplifying RNA Targets.” Physically, it is essentially a series of tubes, with bulbs on the ends of each, etched like channels into the biochip.

There are other pathogen-diagnostic detectors, notably the Polymerase Chain Reaction device (which targets DNA) and the Nucleic Acid Sequence Based Amplification (which also targets RNA). The SMART detector is unique in that the engineers use a DNA probe with base letters that match the code in the targeted sequence. This ensures the probe will latch on only to the specific RNA strand being assayed. The team inundates the sample with probes, to ensure that all RNA molecules bind to a probe.

“The device allows us to design probes that are both sensitive and specific,” Tripathi says.

This approach creates excess—that is, probes with no RNA partners. That’s OK, because the Brown-led team then attached the probes to 2.8 micron magnetic beads that carry the genetic sequence for the influenza RNA sequence.

The engineers then use a magnet to slowly drag the RNA-probe pairs collected in the bulb through a tube that narrows to 50 microns and then deposit the probes at a bulb at the other end.

This convergence of magnetism (the magnetized probes and the dragging magnets) and microfluidics (the probes’ movement through the narrowing channel and the bulbs) serves to separate the RNA-probe pairs from the surrounding biological debris, allowing clinicians to isolate the influenza strains readily and rapidly for analysis. The team reports that it tracks the RNA-probe beads flawlessly at speeds up to 0.75 millimeters per second.

“When we amplify the probes, we have disease detection,” Tripathi says. “If there is no influenza, there will be no probes (at the end bulb). This separation part is crucial.”

Once separated, or amplified, the RNA can be analyzed using conventional techniques, such as nucleic acid sequence-based amplification (NASBA).

The chips created in Tripathi’s lab are less than two inches across, can fit four tube-and-bulb channels, and could be commercially manufactured and made so more channels could be etched on each.

The team is working on separate technologies for biohazard detection.

Stephanie McCalla, who earned her doctorate at Brown last year and is now at the California Institute of Technology (Caltech), is the first author of the paper. The U.S. National Institutes of Health and the National Science Foundation funded the research.

Pictured at the top is Anubhav Tripathi (Credit: Mike Cohea).

Tuesday, June 12, 2012

Rapid Methods Community Mourns the Loss of Dr. Thomas Montag

Dr. Sven M. Deutschmann and Axel H. Schroeder, Chairman and Administration Manager of the ECA Rapid Methods Working Group, respectively, today informed the RMM community of the passing of Dr. Thomas Montag-Lessing, on Friday, May 18.

Dr. Montag-Lessing was deeply involved in the foundation of the European Compliance Academy's Rapid Microbiological Methods Working Group in 2006 and was – to the end – an active member of the group's Advisory Board. As part of his commitment to the European Compliance Academy, he brought his experience and expertise as a speaker and moderator in many microbiological and pyrogen conferences.

Dr. Montag-Lessing studied Medicine at the Humboldt University in Berlin and specialized in Medicinal Microbiology. After spending some years in research work at the Charité in Berlin, he had been employed at the Paul-Ehrlich-Institute since 1990 where he led the Department for Parasitology, Diagnostics and Microbial Safety.

Moreover, he was actively involved in different pharmacopoeial working groups for many years and pushed forward the development of new microbiological methods thanks to his comprehensive expertise – e.g. in the field of endotoxin and pyrogen testing in Germany and Europe.

The Group and all its members will miss him and his innovative contributions and ideas. His death leaves a void that will be hard to fill. In the name of the ECA RMM Working Group and the Advisory Board, we express our profound sadness to his family and relatives.

In memorandum of Dr. Montag-Lessing, I respectfully share the following photograph, which was taken during the initial founding of the ECA RMM Working Group in 2006. Pictured from left to right are Dr. Paul Newby, GlaxoSmithKline; Dr. Christina Bruntner, Pall Life Sciences; Dr. Robert Johnson, Dialogue; Dr. Thomas Lukow, Novartis Behring; Elisabeth Jander, Pall Life Sciences; Mike Edgington, Director for Regulatory Affairs (ECA); Dr. Thomas Mikosch, Centocor; Dr. Klaus Haberer, Compliance Advice and Services in Microbiological; Dr. Thomas Montag, Paul-Ehrlich-Institut; Dr. Sven Deutschmann, Roche Diagnostics; Dr. Ulrich Herber, Concept Heidelberg.

Monday, June 4, 2012

Our LinkedIn Group: Over 2000 Members!

Today our LinkedIn Group, Rapid Micro Methods, surpassed the 2000 member mark! Come join us for discussions about rapid methods that impact a variety of industry, consumer and clinical sectors. Membership in LinkedIn is FREE, and you must have a LinkedIn account to join our discussion forum. You can visit our group by Clicking Here.

The group's members are from a diverse number of companies, academic institutions, regulatory agencies and other professional organizations, all interested in learning about and promoting the implementation of rapid microbiological methods.

STEC Office Building, Tokyo

In consideration of today's new requirement for testing U.S. beef trimmings for six Shiga toxin-producing Escherichia coli (STEC), known as non-O157 Shiga-toxin producing E. coli (STECs) or “The Big Six," I thought it appropriate to share an interesting photo I took during a recent visit to Tokyo. The STEC Jyoho building, located in the middle of the city, is a skyscraper that is very close to the Shinjuku rail station, and houses a number of high profile businesses. Interestingly, one of the building's occupants is the Mitsukoshi Health and Welfare Foundation. 

Expanded E.coli Testing Of U.S. Beef Starts Today

USDA’s Food Safety and Inspection Service (FSIS) will begin routine sampling for six Shiga toxin-producing Escherichia coli (STEC), known as non-O157 Shiga-toxin producing E. coli (STECs) or “The Big Six, in addition to E. coli O157:H7, in raw beef trimmings beginning today.

As part of its a zero-tolerance policy for E. coli O157:H7, FSIS will initially sample raw beef manufacturing trimmings and other raw ground beef product components produced domestically and imported, and test the samples for the serogroups O26, O103, O45, O111, O121 and O145. The Centers for Disease Control and Prevention (CDC) identified these particular serogroups of non-O157:H7 Shiga-toxin producing E.coli, or non-O157 STEC, as those responsible for the greatest numbers of non-O157 STEC illnesses, hospitalizations, and deaths in the United States.

The agency extended the original March implementation date to provide additional time for establishments to validate their test methods and detect the pathogens prior to entering the stream of commerce.

The notice was published on May 31, 2012 in the U.S. Federal Register and includes responses to the comment period that was opened Sept. 20, 2011.

E.coli O157:H7 was declared an adulterant in 1994, following the Jack in the Box E.coli outbreak that sickened hundreds of people and killed four including a young boy. But E.coli O157:H7 is not the only strain of the bacteria that can cause serious illness or death. The U.S. Department of Agriculture’s (USDA) Food Safety and Inspection Service (FSIS) estimates that banning the six additional strains of E.coli will reduce by 110,000 the number of foodborne illnesses reported in the U.S. each year.

Any beef trim testing positive for these pathogens will not be allowed into commerce and will be subject to recall.

“These strains of E. coli are an emerging threat to human health and the steps we are taking today are entirely focused on preventing Americans from suffering foodborne illnesses,” said Agriculture Secretary Tom Vilsack, in a statement. “We cannot ignore the evidence that these pathogens are a threat in our nation’s food supply.”

A number of rapid methods designed to detect these strains are available and can certainly play a role in monitoring the food supply accordingly.

Friday, June 1, 2012

Biochip-based Device for Cell Analysis

Inexpensive, portable devices that can rapidly screen cells for leukemia, HIV and other pathogens may soon be possible thanks to a chip that can produce three-dimensional focusing of a stream of cells, according to researchers.

"HIV is diagnosed based on counting CD4 cells," said Tony Jun Huang, associate professor of engineering science and mechanics, Penn State. "Ninety percent of the diagnoses are done using flow cytometry."

Huang and his colleagues designed a mass-producible device that can focus particles or cells in a single stream and performs three different optical assessments for each cell. They believe the device represents a major step toward low-cost flow cytometry chips for clinical diagnosis in hospitals, clinics and in the field.

"The full potential of flow cytometry as a clinical diagnostic tool has yet to be realized and is still in a process of continuous and rapid development," the team said in a recent issue of Biomicrofluidics. "Its current high cost, bulky size, mechanical complexity and need for highly trained personnel have limited the utility of this technique."

Flow cytometry typically looks at cells in three ways using optical sensors. Flow cytometers use a tightly focused laser light to illuminate focused cells and to produce three optical signals from each cell. These signals are fluorescence from antibodies bound to cells, which reveals the biochemical characteristics of cells; forward scattering, which provides the cell size and its refractive index; and side scattering, which provides cellular granularity. Processingthese signals allows diagnosticians to identify individual cells in a mixedcell population, identify fluorescent markers and count cells and other analysis to diagnose and track the progression of HIV, cancer and other diseases.

"Current machines are very expensive costing $100,000," said Huang. "Using our innovations, we can develop a small one that could cost about $1,000."

One reason the current machines are so large and expensive is the method used to channel cells into single file and the necessary alignment of lasers and multiple sensors with the single-file cell stream. Currently, cells are guided into single file using a delicate three-dimensional flow cell that is difficult to manufacture. More problematic is that these current machines need multiple lenses and mirrors for optical alignment.
"Our approach needs only a simple one-layer, two-dimensional flow cell and no optical alignment is required," said Huang.

Huang and his team used a proprietary technology named microfluidic drifting to create a focused stream of particles. Using a curved microchannel, the researchers took advantage of the same forces that try to move passengers in a car to the outside of a curve when driving. The microfluidic chip's channelbegins as a main channel that contains the flow of carrier liquid and a second channel that comes in perpendicularly that carries the particles or cells. Immediately after these two channels join, the channel curves 90 degrees, which moves all the cells into a horizontal line. After the curve, liquid comes into the channel on both sides, forcing the horizontal line of cells into single file. The cells then pass through a microlaser beam.

An advantage of this microfluidic flow cytometry chip is that it can be mass-produced by molding and standard lithographic processes. The fibers for the optical-fiber delivered laser beams and optical signals already exist.

"The optical fibers are automatically aligned once inserted into the chip, therefore requiring no bulky lenses and mirrors for optical alignment," said Huang. "Our machine is small enough it can be operated by battery, which makes it usable in Africa and other remote locations."

The researchers tested the device using commercially available, cell-sized fluorescent beads. They are now testing the device with actual cells.