MassTag-PCR

MassTag PCR is a modification of PCR based on mass-spectrometric detection of an end product. This technology was developed by researchers from the Mailman School of Public Health at Columbia University and the Columbia Genome Center.[1][2]

Principles

Like conventional PCR, MassTag-PCR uses primer pairs. The difference is primers used for MassTag PCR are tagged with molecules of known masses or MassCodes. Instead of single pair of primers this technology uses a number of primers, making it a multiplex system. Unlike, conventional Multiplex PCR system, in MassTag PCR more than 15 primer pairs could be used. If DNA from any of the agent of primer panel is present, it will be amplified. Each amplified product will carry its specific Masscodes. The PCR product is then purified to remove unbound primers, dNTPs, enzyme and other impurities. Finally, the purified PCR products are subjected to UV light as the chemical bond between the nucleic acids and primers is photolabile. As the Masscodes are liberated from PCR products they are detected with a Mass Spectrometer. Presence of specific MassCode indicates the presence of specific pathogen.[1]

Applications

This technique has been found to detect a previously uncharacterized clade of rhinovirus.[3] MassTag PCR is a more comprehensive and sensitive diagnostic technique, CII was able to determine the cause of this illness for 26 out of 79 previously unknown cases. MassTag PCR demonstrated its tripartite value as a tool for surveillance, outbreak detection, and epidemiology.

References

  1. ^ a b Briese, Thomas; Palacios, Gustavo; Kokoris, Mark; Jabado, Omar; Liu, Zhiqiang; Renwick, Neil; Kapoor, Vishal; Casas, Inmaculada; Pozo, Francisco; Limberger, Ron; Perez-Brena, Pilar; Ju, Jingyue; Lipkin, W. Ian (2005). "Diagnostic System for Rapid and Sensitive Differential Detection of Pathogens". Emerging Infectious Diseases. 11 (2): 310–313. doi:10.3201/eid1102.040492. ISSN 1080-6040. PMC 3320438. PMID 15752453.
  2. ^ Larkin, Marilynn (2005). "Technology and public health". The Lancet Infectious Diseases. 5 (9): 542. doi:10.1016/s1473-3099(05)70211-5. ISSN 1473-3099.
  3. ^ Dominguez, Samuel R.; et al. (October 2008). "Multiplex MassTag-PCR for respiratory pathogens in pediatric nasopharyngeal washes negative by conventional diagnostic testing shows a high prevalence of viruses belonging to a newly recognized rhinovirus clade". Journal of Clinical Virology. 43 (2): 219–222. doi:10.1016/j.jcv.2008.06.007. PMC 2603178. PMID 18674964.

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