Rapid Identification of Biothreat and Other Clinically Relevant Bacterial Species by Use of Universal PCR Coupled with High-Resolution Melting Analysis

ABSTRACT A rapid assay for eubacterial species identification is described using high-resolution melt analysis to characterize PCR products. Unique melt profiles generated from multiple hypervariable regions of the 16S rRNA gene for 100 clinically relevant bacterial pathogens, including category A and B biothreat agents and their surrogates, allowed highly specific species identification.

[1]  Paul Matsudaira,et al.  A 768-lane microfabricated system for high-throughput DNA sequencing. , 2005, Lab on a chip.

[2]  L. Rotz,et al.  Advances in detecting and responding to threats from bioterrorism and emerging infectious disease , 2004, Nature Medicine.

[3]  F. Burkle Mass casualty management of a large-scale bioterrorist event: an epidemiological approach that shapes triage decisions. , 2002, Emergency medicine clinics of North America.

[4]  Justin Hardick,et al.  Rapid PCR-Based Diagnosis of Septic Arthritis by Early Gram-Type Classification and Pathogen Identification , 2008, Journal of Clinical Microbiology.

[5]  Carl T Wittwer,et al.  High-resolution genotyping by amplicon melting analysis using LCGreen. , 2003, Clinical chemistry.

[6]  Samuel Yang,et al.  PCR-based diagnostics for infectious diseases: uses, limitations, and future applications in acute-care settings , 2004, The Lancet Infectious Diseases.

[7]  D. Alland,et al.  A detailed analysis of 16S ribosomal RNA gene segments for the diagnosis of pathogenic bacteria. , 2007, Journal of microbiological methods.

[8]  B. Robertson,et al.  New microbiology tools for public health and their implications. , 2005, Annual review of public health.

[9]  C. Tseng,et al.  Rapid detection and identification of clinically important bacteria by high-resolution melting analysis after broad-range ribosomal RNA real-time PCR. , 2006, Clinical chemistry.

[10]  Thomas C. Quinn,et al.  Quantitative Multiprobe PCR Assay for Simultaneous Detection and Identification to Species Level of Bacterial Pathogens , 2002, Journal of Clinical Microbiology.

[11]  J. Clarridge,et al.  Impact of 16S rRNA Gene Sequence Analysis for Identification of Bacteria on Clinical Microbiology and Infectious Diseases , 2004, Clinical Microbiology Reviews.

[12]  Rangarajan Sampath,et al.  Rapid identification and strain-typing of respiratory pathogens for epidemic surveillance , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[13]  D. Lim,et al.  Current and Developing Technologies for Monitoring Agents of Bioterrorism and Biowarfare , 2005, Clinical Microbiology Reviews.

[14]  Justin Hardick,et al.  Rapid polymerase chain reaction-based screening assay for bacterial biothreat agents. , 2008, Academic emergency medicine : official journal of the Society for Academic Emergency Medicine.