title: Comparative Genomic Analysis of Haemophilus haemolyticus and Nontypeable Haemophilus influenzae and a New Testing Scheme for Their Discrimination.
abstract: Haemophilus haemolyticus has been recently discovered to have the potential to cause invasive disease. It is closely related to nontypeable Haemophilus influenzae (NT H. influenzae). NT H. influenzae and H. haemolyticus are often misidentified because none of the existing tests targeting the known phenotypes of H. haemolyticus are able to specifically identify H. haemolyticus Through comparative genomic analysis of H. haemolyticus and NT H. influenzae, we identified genes unique to H. haemolyticus that can be used as targets for the identification of H. haemolyticus A real-time PCR targeting purT (encoding phosphoribosylglycinamide formyltransferase 2 in the purine synthesis pathway) was developed and evaluated. The lower limit of detection was 40 genomes/PCR; the sensitivity and specificity in detecting H. haemolyticus were 98.9% and 97%, respectively. To improve the discrimination of H. haemolyticus and NT H. influenzae, a testing scheme combining two targets (H. haemolyticus purT and H. influenzae hpd, encoding protein D lipoprotein) was also evaluated and showed 96.7% sensitivity and 98.2% specificity for the identification of H. haemolyticus and 92.8% sensitivity and 100% specificity for the identification of H. influenzae, respectively. The dual-target testing scheme can be used for the diagnosis and surveillance of infection and disease caused by H. haemolyticus and NT H. influenzae.
authors.affiliation: Meningitis and Vaccine Preventable Diseases Branch, Division of Bacterial Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors.forename: Fang
authors.initials: F
authors.lastname: Hu
authors.affiliation: School of Biology, Georgia Institute of Technology, Atlanta, Georgia, USA.|Applied Bioinformatics Laboratory, Atlanta, Georgia, USA.|PanAmerican Bioinformatics Institute, Cali, Valle del Cauca, Colombia.
authors.forename: Lavanya
authors.initials: L
authors.lastname: Rishishwar
authors.affiliation: School of Biology, Georgia Institute of Technology, Atlanta, Georgia, USA.
authors.forename: Ambily
authors.initials: A
authors.lastname: Sivadas
authors.affiliation: School of Biology, Georgia Institute of Technology, Atlanta, Georgia, USA.|Institute of Science and Technology Austria, Klosterneuburg, Austria.
authors.forename: Gabriel J
authors.initials: GJ
authors.lastname: Mitchell
authors.affiliation: School of Biology, Georgia Institute of Technology, Atlanta, Georgia, USA.|Applied Bioinformatics Laboratory, Atlanta, Georgia, USA.|PanAmerican Bioinformatics Institute, Cali, Valle del Cauca, Colombia.
authors.forename: I King
authors.initials: IK
authors.lastname: Jordan
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Clinical and Translational Research Center, Department of Microbiology and Immunology, University at Buffalo, State University of New York, Buffalo, New York, USA.
authors.forename: Timothy F
authors.initials: TF
authors.lastname: Murphy
authors.affiliation: Department of Pediatrics, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Janet R
authors.initials: JR
authors.lastname: Gilsdorf
authors.affiliation: Meningitis and Vaccine Preventable Diseases Branch, Division of Bacterial Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors.forename: Leonard W
authors.initials: LW
authors.lastname: Mayer
authors.affiliation: Meningitis and Vaccine Preventable Diseases Branch, Division of Bacterial Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA gqe8@cdc.gov.
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
mesh_terms: D001483:Base Sequence; D055028:Comparative Genomic Hybridization / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics; D016680:Genome, Bacterial / Q000235:genetics; D006192:Haemophilus Infections / Q000175:diagnosis* / Q000382:microbiology; D006193:Haemophilus influenzae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D057230:Limit of Detection; D008074:Lipoproteins / Q000235:genetics*; D051026:Phosphoribosylglycinamide Formyltransferase / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial; D008074:Lipoproteins; D051026:Phosphoribosylglycinamide Formyltransferase