Duplex Quantitative PCR Assay for Detection of Haemophilus influenzae That Distinguishes Fucose- and Protein D-Negative Strains

Camilla de Gier; Janessa L Pickering; Peter C Richmond; Ruth B Thornton; Lea-Ann S Kirkham

doi:10.1128/JCM.00982-16

Duplex Quantitative PCR Assay for Detection of Haemophilus influenzae That Distinguishes Fucose- and Protein D-Negative Strains

J Clin Microbiol. 2016 Sep;54(9):2380-3. doi: 10.1128/JCM.00982-16. Epub 2016 Jun 22.

Authors

Camilla de Gier¹, Janessa L Pickering², Peter C Richmond³, Ruth B Thornton², Lea-Ann S Kirkham⁴

Affiliations

¹ School of Paediatrics and Child Health, University of Western Australia, Perth, Australia.
² School of Paediatrics and Child Health, University of Western Australia, Perth, Australia Wesfarmers Centre for Vaccines and Infectious Diseases, Telethon Kids Institute, Perth, Australia.
³ School of Paediatrics and Child Health, University of Western Australia, Perth, Australia Wesfarmers Centre for Vaccines and Infectious Diseases, Telethon Kids Institute, Perth, Australia Department of Paediatrics, Princess Margaret Hospital for Children, Subiaco, Australia.
⁴ School of Paediatrics and Child Health, University of Western Australia, Perth, Australia Wesfarmers Centre for Vaccines and Infectious Diseases, Telethon Kids Institute, Perth, Australia lea-ann.kirkham@uwa.edu.au.

Abstract

We have developed a specific Haemophilus influenzae quantitative PCR (qPCR) that also identifies fucose-negative and protein D-negative strains. Analysis of 100 H. influenzae isolates, 28 Haemophilus haemolyticus isolates, and 14 other bacterial species revealed 100% sensitivity (95% confidence interval [CI], 96% to 100%) and 100% specificity (95% CI, 92% to 100%) for this assay. The evaluation of 80 clinical specimens demonstrated a strong correlation between semiquantitative culture and the qPCR (P < 0.001).

MeSH terms

Bacterial Proteins
Carrier Proteins
Fucose / deficiency*
Haemophilus influenzae / genetics*
Haemophilus influenzae / isolation & purification*
Humans
Immunoglobulin D / deficiency*
Lipoproteins / deficiency*
Multiplex Polymerase Chain Reaction / methods*
Real-Time Polymerase Chain Reaction / methods*
Sensitivity and Specificity

Substances

Bacterial Proteins
Carrier Proteins
Immunoglobulin D
Lipoproteins
Fucose
glpQ protein, Haemophilus influenzae