Medical Device Link.

Originally Published IVD Technology April 2003

Beyond Clinical Diagnostics

While the government is involved in developing biodefense diagnostics, it needs better coordination.

by Richard Park

The world has become a different place since September 11. Two years ago, the average person did not know much about bioterrorism, biowarfare, and biodefense. Today, most everyone has become well aware of what these issues are, to the extent that they have affected how people carry out their daily lives.

The threat of bioterrorism also presents new challenges to the approach of government agencies toward security matters. Since bioterrorism presents a different type of threat that could rapidly cause widespread harm, the government has taken an active role in finding ways to deal with this threat. For example, the government has looked into what needs to be done in the event of a bioterrorist attack, including how to detect whether an attack has occurred and how to provide treatment to victims. 

However, the government realizes fighting bioterrorism is a massive endeavor, one it cannot handle alone. In particular, developing biodefense mechanisms requires an immense investment of technical expertise. Even though the government itself does have considerable scientific knowledge at its disposal, it has turned to the business and academic communities and solicited their help in the biodefense arena. 

One important area in biodefense is developing technologies that can identify and test for the presence of biothreat agents, such as anthrax, smallpox, and sarin. The government has been working with IVD companies to develop such technologies, utilizing their knowledge to produce diagnostic systems for biodefense purposes. While these efforts have resulted so far in effective technologies that are currently in use, the government plans to continue developing and improving biodefense diagnostics and getting the IVD industry involved in this effort. 

Government Activity

The autonomous pathogen detection system (APDS) by Lawrence Livermore National Laboratory (Livermore, CA) is a continuous and fully automated monitoring system that can identify pathogens in the air.

Since bioterrorism affects all walks of life and all areas of public safety, nearly every government agency has been involved to some degree in developing biodefense diagnostics. Whether it is CDC, EPA, or FDA, the involvement of such agencies has ranged from pledging financial resources to conducting the actual research for diagnostic technologies. The activities of government agencies that have played a significant role in developing such technologies are discussed below.

NIAID. Since the fall of 2001, the National Institute of Allergy and Infectious Diseases (NIAID; Bethesda, MD), a division of the National Institutes of Health, has accelerated its biodefense research program by launching more than 30 new research initiatives. NIAID has also created a comprehensive strategic plan and a detailed research agenda for category A biothreat agents, those that CDC has identified as being most dangerous. Cateogry A agents include the viruses that cause smallpox and hemorrhagic, or bleeding, fevers such as ebola. These agents also include the bacteria that cause anthrax, the plague, botulism, and tularemia. 

For each organism that might be used as a biothreat agent, NIAID has set priorities and goals in a number of different research categories. One of these categories is diagnostics. In order to support the development of biodefense diagnostics, NIAID has set up various research grant programs. As reported in the March 2003 issue of IVD Technology (Notable, p. 15), one program that may be of particular interest to IVD companies is the biodefense partnerships that are geared toward facilitating collaboration between government, academia, and the private sector. 

“NIAID right now is supporting research and development in technologies with the ultimate goal of supporting diagnostics,” says Maria Giovanni, PhD, assistant director for microbial genomics and advanced technologies at NIAID. “We have programs in place that can engage industry to partner with NIAID to develop these technologies. And they range from very early development of a technology to later-stage development.”

The NIAID grant program supports the further development of sensitive, specific, and rapid diagnostic methods and platforms that can distinguish biothreat pathogens from common infections. NIAID anticipates that applications for these biodefense partnership grants will focus on technologies that have previously shown promise in the early stages of development. 

The RAMP reader by Response Biomedical Corp. (Burnaby, BC, Canada) can perform tests to detect various biowarfare agents, such as anthrax and ricin.

In addition, NIAID has placed a high priority on these biodefense diagnostics and encourages applicants to focus on the following areas:

• Tests that evaluate the possible spectrum of antimicrobial resistance or genetic manipulation.
• Tests capable of high-throughput screening (e.g., microchip-based platforms) containing microbial signature profiles.
• Tests capable of identifying multiple pathogens simultaneously in a single sample.
• Novel assays based on human immune or other physiological responses.
• Tests capable of identifying novel biomarkers for human immune activation.
• In vivo imaging methods and development of contrast reagents for visualization of pathogens or host immune responses in vivo.
• Clinical diagnostic tools for human eczema.
• Diagnostics that can be used in remote settings. 

“The partnership grant actually has to have a significant industry partner,” says Giovanni. “So it really is geared toward industry. If there’s not a substantial industry component, it’s not a partnership grant.”

The deadline for biodefense partnership grant applications is June 10, 2003. Additional information about these research grant programs can be accessed through NIAID’s Web site at http://www.niaid.nih.gov/biodefense.

USAMRIID. The U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID; Fort Detrick, MD) is the lead medical research laboratory for the U.S. Biological Defense Research Program. Its mission is to provide medical countermeasures to biological weapons. USAMRIID is divided into four main research divisions, one of which is devoted solely to diagnostics. All of the institute’s development work and much of the basic research that is required to develop diagnostics are done within its diagnostics division. 

This division itself is also divided into several different branches. One branch focuses its research on molecular diagnostics, such as nucleic acid detection methods, polymerase chain reaction (PCR), and other related technologies. Another branch develops antigen and antibody detection methods as well as immunodiagnostics. 

Another branch of the diagnostics division is responsible for training and field operations. Since the diagnostics that are developed eventually get transitioned out and the users in the military need to learn how to conduct these tests, this branch trains users to perform and interpret the tests. It is also charged with developing plans to test the diagnostics in the field environment and to make sure that the tests function the same way in difficult climates and conditions, similar to what would be expected in a military deployment situation. 

One other branch that has been combined with the field operations and training branch is the special pathogen sample testing laboratory. It is responsible for interfacing with the Department of Defense and other government agencies to support operational missions by testing samples that may come in for analysis. 

The handheld advanced nucleic acid analyzer (HANAA) by Lawrence Livermore National Laboratory is a portable, PCR-based instrument that can identify pathogens based on their DNA.

“We’re responsible not only for developing diagnostics, but also for actually doing a lot of the diagnostics themselves,” says George Ludwig, PhD, chief of the diagnostic systems at USAMRIID. “And in some cases, we conduct environmental testing in an operations scenario.”

According to Ludwig, USAMRIID’s diagnostic division has been working on a wide array of biodefense technologies. During the last 5–10 years, their research has focused on bringing into field use second-generation diagnostic technologies, primarily advanced immunodiagnostics and nucleic acid–based detection methods.

“We’ve worked very closely with commercial companies to look at platforms the government needs,” says Ludwig. “We’ve taken the platforms that do best at meeting our needs and developed assays for those. Now we’re at the point where we’re putting those technologies into the hands of the field-deployed units, and they’re being used. So one component of our research effort is focused on supporting those technologies. We’re supporting those technologies with improved assays and by continuing to upgrade their capabilities.” 

At the same time, USAMRIID is already looking beyond, toward third-generation diagnostics. For such diagnostics, the institute plans to take the same approach as it has in the past: collaborating with industrial partners and other government organizations to look at what new platforms can be brought forward.

“Essentially what we’d like to have some time in the distant future is a tricorder type of device that can give 100%-confident results to a diagnostic test and can test multiple biomarkers in a very rapid fashion,” says Ludwig.

LLNL. Lawrence Livermore National Laboratory (LLNL; Livermore, CA) is a Department of Energy national laboratory operated by the University of California. LLNL has set up a number of homeland security programs, including the chemical and biological national security program. This program is addressing the need for technologies that can quickly detect, identify, and mitigate the use of chemical and biological threat agents. 

“The central theme of our diagnostic devices is can we detect something earlier because our primary focus has been on civilian counterterrorism prep applications,” says Richard Langlois, PhD, chief scientist of the chemical and biological national security program at LLNL. “In a situation like that, oftentimes people may not even notice anything has happened. So being able to detect things as soon as possible to let people take countermeasures is important.”

One of the diagnostic technologies that LLNL has already developed and put into use is the biological aerosol sentry and information system (BASIS). This system consists of a network of air sampling units that collect and check air particles. The filters in these units capture the air particles and are then collected for analysis several times a day. A transportable field laboratory analyzes the samples using sensitive DNA-based identification techniques that have been validated by CDC. BASIS was part of the security network deployed during the Salt Lake City Winter Olympics in 2002.

“The idea behind BASIS was: could we take off-the-shelf technology right now, and with some minor modifications, get it ready to go and be used in the field,” says Langlois. “So the basic approach was to take large commercial air samplers that would suck in air and deposit particles on filters, and then use commercial PCR instruments that do the DNA analysis of those filters and test for pathogens.” 

As a follow-up to BASIS, LLNL has also developed the autonomous pathogen detection system (APDS). This system is a continuous and fully automated monitoring system that identifies pathogens that may be in the air. The size of a mailbox, APDS has an air collector, a fluidic module that mixes the samples and reagents and performs all the biochemistry, and a detection module. Unlike BASIS, for which personnel have to collect the samples and take them to a lab, APDS is a fully self-contained system that will continually conduct tests and report when it detects something. If a pathogen is detected, the information is sent via a network to a central location for further analysis and response.

LLNL has assembled some prototypes of APDS and is in the process of licensing it to a manufacturer. LLNL is hoping to have this system commercially available in a year. 

Another biodefense diagnostic that LLNL is developing is the handheld advanced nucleic acid analyzer (HANAA). This device is a portable, PCR-based instrument that can identify pathogens based on their DNA. The size of a brick, HANAA has four different PCR chambers in which analyses can be conducted simultaneously. Many cycles of heating and cooling can be done very rapidly within the analyzer, so a full PCR can be done in 15–30 minutes. Since it is such a miniaturized device, HANAA is intended to be taken out and used in the field to run tests for biothreat agents.
“So a member of a haz-mat team can take this device right up to the site of a spill or a suspect powder, set up a reaction, and do the test right on the spot,” says Langlois.

Developing New Diagnostics

Despite the programs that have been put in place and the technologies that either have been developed or are currently being developed, government agencies know that their work in fighting bioterrorism has only just begun. Terrorists will produce new strains of biothreat agents, so the onus is on the government and IVD companies to continue doing research and improving biodefense diagnostic tools.

The GeneXpert by Cepheid Inc. (Sunnyvale, CA) is a compact system with four independent PCR-reactor modules and disposable single-test cartridges.

One area the government is particularly focusing on is developing an integrated diagnostic platform. Such a technology would allow a number of different tests, such as nucleic acid and antigen detection and host response monitoring, to be performed on a single platform. The technologies that researchers have been considering and investing money into toward developing an integrated platform are kit-based technologies and microarrays, since both offer the ability to conduct multiple tests in a single platform. Government officials acknowledge that the main reason for this emphasis on developing an integrated diagnostic technology stems from lessons that were learned following September 11.

“We learned that multiple technologies and looking at multiple biomarkers can provide very high confidence to answers and test results,” says Ludwig. “The problem is how to put that all together in a platform that doesn’t really take a whole lot of time. The technology to do that is being developed. It’s not there yet, but it’s coming.”

In trying to decide which IVD companies to work with in developing an integrated diagnostic platform, officials said it relates to the strategic idea of an integrated system. According to officials, many companies have good immunodiagnostic tests that are capable of detecting bacteria. However, that answers only one small part for an integrated system. The government is more likely to partner with a company that has the same kind of strategic plan and is working toward developing an integrated system that can do everything at one time. The government also considers several other factors.

“We take into account the performance characteristics of a company’s system, such as sensitivity, and specificity,” says Ludwig. “Speed is another factor, how fast the assays can be done. If there’s no benefit in the system over what I’m currently using, it doesn’t make much sense to partner with that company. So I use a series of selection criteria that helps me limit the number of people that we’re likely to partner with.”

Other officials believe that more of the biodefense diagnostic research effort will go toward increasing the range of diagnostics available. Since there are still a lot of viruses for which no good assays exist, researchers hope to broaden the scope of assay development for biothreat agents. The government is also looking into developing instrumentation that can process large numbers of samples simultaneously. If such an instrument could be developed so that every biothreat sample collected could be tested for a variety of pathogens, researchers might be able to pick up something that had not yet been diagnosed through other methods. 

In addition, the government plans to continue trying to understand better the mechanisms of pathogenesis. Researchers want to know how the host responds to an infection, what are the potential markers, and what more can be learned about how this process occurs so that bioterrorist attacks can be detected earlier. 

“There are a number of emerging diseases that have come out in the last 10–20 years that have been surprising to us,” said Langlois. “We continue to work on mechanisms to find these things and detect them, whether they’re natural or man-made.”

Coordinating Government Efforts

The force amplified biological sensor (FABS) by Purdue University (West Lafayette, IN) is a rapid, automated assay that can detect analytes such as proteins, viruses, and bacteria.

During the past two and a half years, there has been a flurry of activity in not only the biodefense diagnostics area but also in the biodefense arena as a whole. Government agencies and IVD companies have been actively involved in conducting research and developing products, and have consequently produced a number of new biodefense technologies.

However, because of all this activity, a considerable amount of confusion has emerged as to which diagnostics have already been developed, which ones are available, and whether or not what has been produced addresses what is needed most. Moreover, there seems to be a lack of leadership on the government’s part in terms of coordinating these agencies and companies toward having a focused effort in developing technologies that can effectively address the most urgent needs. Government officials recognize this problem and have been working on fixing this problem.

“All of those government agencies that are getting into the biodefense area have different goals and requirements,” says Ludwig. “We recognized that almost immediately after September 11 when a lot of these government agencies started jumping on the bandwagon. What we’re doing now is trying to get organized and figure out where we can make better progress in the area of diagnostics. This has only really blossomed in the last year and a half, and during that period of time, a lot of government agencies have been getting together both formally and informally. And that is expanding. We’re actually beginning to make some very significant progress in the sharing of information.”

Moreover, officials are hoping that the recently formed Department of Homeland Security (DHS) will take a lead role in streamlining the process and formulating strategy for developing biodefense technologies. 

“The goal of DHS is to bring together all of these entities, not only specifically to help bring the collective minds of technology, but also to bring the collective resources of all the agencies together into one organization,” says Brian Roehrkasse, DHS spokesman. “This is an exciting time because for the first time, the federal government has one concerted effort to work with a partnership to foster the development of technologies that will protect the homeland. And through our Homeland Security Advanced Research Project Agency, we will have the opportunity to work with the private sector to develop these technologies.”

IVD industry analysts agree that while the government has been disjointed and disorganized in its efforts to develop biodefense diagnostics, DHS should be able to clarify some of the confusion and provide guidance in the future.
“One flaw right now is that we don’t have the system coordinated among the various government agencies as well as it should be,” says Tom Gutshall, chairman of the board at Cepheid (Sunnyvale, CA). “That’s what DHS is trying to sort out. It was obvious to us very early on that it was a disjointed effort, it didn’t seem to have a leader, no czar, no one who was overseeing and directing how one should go about mounting the defense. The focus now on DHS is going to bring a lot of these agencies into a much closer working relationship with each other. We should begin to see efficiencies as this proceeds further, and we’ve seen some indications of that already.”

Conclusion

Ever since September 11 and the anthrax attacks that occurred the following month, the public has become increasingly skittish about a possible bioterrorist attack. While in the past high-profile events like the Olympics, the Super Bowl, and the Academy Awards were opportunities to celebrate human achievement, they are now marred by concerns over bioterrorism and making preparations in the event of an attack.

Recent elevations of the national threat level to a high risk of terrorist attack, or level orange, have sent many people rushing to their local hardware stores to stock up on plastic sheeting and duct tape. It is a sad state of affairs, but unfortunately the reality of the current times. What is hoped is that the government and the IVD industry will continue working closely to develop effective biodefense diagnostic technologies.

Richard Park is editor of IVD Technology. 

Copyright ©2003 IVD Technology