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13:56
A Comparative Approach to Characterize the Landscape of Host-Pathogen Protein-Protein Interactions
Published on: July 18, 2013
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Haemophilus influenzae: using comparative genomics to accurately identify a highly recombinogenic human pathogen
Erin P Price1,2, Derek S Sarovich3, Elizabeth Nosworthy4
1Child Health Division, Menzies School of Health Research, Darwin, NT, Australia. erin.price@menzies.edu.au.
BMC Genomics
|August 28, 2015
Summary
Accurate identification of Haemophilus influenzae is challenging due to species similarity. This study developed a highly specific fucP real-time PCR assay for reliable detection of H. influenzae.
Area of Science:
- Microbiology
- Genomics
- Molecular Diagnostics
Background:
- Haemophilus influenzae is a significant human pathogen causing acute and chronic diseases.
- Accurate identification is difficult due to similarity with H. haemolyticus and genetic exchange within Haemophilus species.
- Existing speciation assays can yield false-positive and false-negative results.
Purpose of the Study:
- To accurately delineate H. influenzae from closely related species using large-scale comparative genomics.
- To identify a species-specific molecular marker for H. influenzae.
- To develop a novel, highly specific diagnostic assay for H. influenzae.
Main Methods:
- Whole-genome single-nucleotide polymorphism (SNP) data from 246 Haemophilus isolates were used for phylogenetic analysis.
- A species-specific locus, fucP, was identified in H. influenzae.
- A TaqMan real-time PCR assay targeting the fucP locus was designed and validated.
Main Results:
- A whole-genome phylogeny successfully differentiated H. influenzae from related species.
- The fucP locus was identified as a unique marker for H. influenzae.
- The novel fucP real-time PCR assay demonstrated 100% specificity for H. influenzae against a panel of clinically relevant species and in silico analysis.
Conclusions:
- This study provides the first large-scale genomic delineation of H. influenzae and identifies fucP as a species-specific marker.
- The fucP assay offers superior accuracy compared to previous H. influenzae targets, many lacking validation in the next-generation genomics era.
- The fucP assay is recommended for accurate clinical and research detection and diagnosis of H. influenzae.
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