From May through October 2009, a total of 10,624 clinical samples from 23 US states were screened for multiple respiratory pathogen gene targets. Of 3,110 (29.3%) samples positive for pandemic (H1N1) 2009 virus, 28% contained ≥1 other pathogen, most commonly Staphylococcus aureus (14.7%), Streptococcus pneumoniae (10.2%), and Haemophilus influenzae (3.5%).
Koon K, Sanders CM, Green J, Malone L, White H, Zayas D, et al. Co-detection of pandemic (H1N1) 2009 virus and other respiratory pathogens. Emerg Infect Dis. 2010 Dec. http://www.cdc.gov/EID/content/16/12/1976.htm
Early treatment of bloodstream infections with appropriate, definitive antimicrobial therapy has proven to reduce mortality, length of hospital stay, and healthcare costs. Culture-based testing methods require up to five days for final pathogen identification and susceptibility reporting, forcing use of broad spectrum empiric therapy.
Edward H. Eiland III, Nicholas Beyda, Jian Han, William Lindgren, RandyWard, Thomas MacAndrew English, Ali Hassoun, and Kathi Hathcock (2010). The Utility of Rapid Microbiological and Molecular Techniques in Optimizing Antimicrobial Therapy. SRX Pharmacology, Volume 2010
This publication reports the use of ResPlex III for genotyping influenza A viruses. The performance characteristics of the assay with regard to H5N1 are further evaluated. The ResPlex system incorporates a novel multiplex PCR technology, target-enriched multiplex PCR, to simultaneously amplify multiple molecular targets in one reaction.
Zou, S., Han, J., Wen, L., Liu, Y., Cronin, K., Lum, S. H., Gao, L., Dong, J., Zhang, Y., Guo, Y., & Shu, Y. (2007). Human Influenza A Virus (H5N1) Detection by a Novel Multiplex PCR Typing Method. Journal of Clinical Microbiology, 45(6), 1889-1892.
While most diagnostic processes cease with the detection of the first relevant infectious agent, newer multiplexed molecular methods which provide simultaneous analysis of multiple agents may give a more accurate representation of the true pathogen spectrum in these samples. To examine this in the context of respiratory infections, acute-phase respiratory specimens submitted to our clinical diagnostic microbiology/virology laboratory for our routine VIRAP diagnosis protocol during the spring 2006 peak respiratory infection season were processed in parallel by analysis with Genaco (QiaPlex) ResPlex I and ResPlex II molecular diagnostic panels.
Brunstein, J.D., Cline, C. L., McKinney, S., & Thomas, E. (2008). Evidence from Multiplex Molecular Assays for Complex Multipathogen Interactions in Acute Respiratory Infections. Journal of Clinical Microbiology, 46(1), 97-102.
This poster describes a study at a large hospital evaluating the usefulness of Tem-PCR in positive blood cultures.
Edward H. Eiland, III., Pharm.D., MBA, BCPS (AQ-ID), Nicholas Beyda, Pharm.D., Jian Han, M.D., Ph.D., Ali Hassoun, M.D., William Lindgren, M.T. (ASCP), Randy Ward, M.T. (ASCP), Thomas M. English, Ph.D.,, and Kathi Hathcock, M(ASCP)SM, CIC
This study reports on the results of a pilot study comparing our clinical diagnostic virology laboratory’s current methods of respiratory pathogen detection against the Genaco Respiratory Infections Panels 1 and 2. These assays employ xMap (Luminex) liquid phase bead conjugated array technology to facilitate automated detection of PCR and RT-PCR products, which provides potential for levels of assay multiplexing above those currently practical with either conventional gel-resolved or real-time methods.
Brunstein, J. & Thomas, E. (2006). Direct Screening of Clinical Specimens for Multiple Respiratory Pathogens Using the Genaco Respiratory Panels 1 and 2. Diagn Mol Pathol, 15(3), 169-173.
Clinical presentations for viral respiratory tract infections are often nonspecific, and a rapid, highthroughput laboratory technique that can detect a panel of common viral pathogens is clinically desirable. We evaluated two multiplex reverse transcription-PCR (RT-PCR) products coupled with microarray-based systems for simultaneous detection of common respiratory tract viral pathogens.
Li, H., McCormac, M. A., Estes, R. W., Sefers, S. E., Dare, R. K., Chappell, J. D., Erdman, D. D., Wright, P. F. & Tang, Y. (2007). Simultaneous Detection and High Throughput Identification of a Panel of RNA Viruses Causing Respiratory Tract Infections. Journal of Clinical Microbiology, 45(7), 2105-2109.
The majority of existing human papillomavirus (HPV) genotyping assays are based on multiplex PCR using consensus or degenerate primers. We developed a Templex HPV assay that simultaneously detects and identifies 25 common HPV genotypes in a single-tube reaction using type-specific primers for the HPV-specific E6 and E7 genes.
Han, J., Swan, D. C., Smith, S. J., Lum, S. H., Sefers, S. E., Unger, E. R., & Tang, Y. (2006). Simultaneous Amplification and Identification of 25 Human Papillomavirus Types with Templex Technology. Journal of Clinical Microbiology, 44(11).