Researchers at Columbia University's Mailman School of Public Health and the Columbia Genome Center have designed and developed a sensitive new diagnostic technology platform, called “Mass Tag PCR,” that can simultaneously screen for multiple infectious agents. The new technology is addressed in a paper published in the February issue of the Centers for Disease Control and Prevention's (CDC) Emerging Infectious Diseases. This new platform is demonstrated in an assay that detects and discriminates 22 pathogens including viruses and bacteria that can present as clinically similar pulmonary disease.
This new technology platform addresses important challenges for infectious disease identification—sensitivity and breadth. Mass Tag PCR provides the ability to be precise in identification, as well as the ability to apply current diagnostics to more than one pathogen at a time, thereby reducing the time needed for differential diagnosis.
“We focused first on respiratory diseases because differential diagnosis is a common clinical problem with implications for outbreak control and individual case management,” stated W. Ian Lipkin, MD, Jerome L. and Dawn Greene Professor of Epidemiology and professor of Neurology and Pathology at Columbia University , and senior author of the paper. “However, we envision implementing this method for a wide variety of applications such as blood product surveillance, agriculture, forensic microbiology, and biodefense.”
To address the need for highly sensitive diagnostics, researchers built on an established method known as polymerase chain reaction that allows amplification of genetic sequences and on a technology previously used for DNA sequencing and detection of genetic polymorphisms. Genetic probes for pathogens were coupled to markers known as mass codes. After amplification, incorporated mass codes were detected by mass spectroscopy allowing identification of the pathogen.
According to Thomas Briese, PhD, associate director of the Greene Infectious Disease Laboratory at the Mailman School, “This new platform can easily detect multiple pathogens in a couple of hours with minimal labor, and will allow comprehensive detection of co-infections.”
“The high multiplexing capability and sensitivity of the approach is provided by using the digital molecular weights of small molecules to code for the identity of the pathogens and detection of the molecular tags with high sensitive mass spectrometry, which can be potentially miniaturized,” stated coauthor, Jingyue Ju, PhD, Head of DNA Sequencing and Chemical Biology at the Columbia Genome Center.
The project, headquartered at the Mailman School 's Jerome L. and Dawn Greene Infectious Disease Laboratory, is a global effort, with partners from around the world providing input into assay design, clinical samples for validation, as well as opportunities to advance the science to the next, practical stages—including outbreak investigation, and blood product screening. Key partners in the work presented in the current publication include the Centro Nacional de Microbiologia of Spain , the Wadsworth Laboratory of the New York State Department of Health, Qiagen Inc, and Operon Inc. Application of the technology is already being evaluated through collaborative relationships with the National Institute for Communicable Diseases, South Africa , the University of Hong Kong , SAR, the Robert Koch-Institute, Germany , and the Bernhard-Nocht-Institute for Tropical Medicine, Germany .
Funding for the project was received from the National Institutes of Health / National Institute of Allergy and Infectious Disease, the Northeast Biodefense Center , and the Ellison Medical Foundation.
Source : Columbia University's Mailman School of Public Health