Coughlan Helena, Reddington Kate, Tuite Nina, Boo Teck Wee, Cormican Martin, Barrett Louise, Smith Terry J, Clancy Eoin, Barry Thomas
Nucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland; Biomedical Diagnostics Institute Programme, National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland.
Nucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
Diagn Microbiol Infect Dis. 2015 Oct;83(2):112-6. doi: 10.1016/j.diagmicrobio.2015.06.013. Epub 2015 Jun 23.
Haemophilus influenzae is recognised as an important human pathogen associated with invasive infections, including bloodstream infection and meningitis. Currently used molecular-based diagnostic assays lack specificity in correctly detecting and identifying H. influenzae. As such, there is a need to develop novel diagnostic assays for the specific identification of H. influenzae. Whole genome comparative analysis was performed to identify putative diagnostic targets, which are unique in nucleotide sequence to H. influenzae. From this analysis, we identified 2H. influenzae putative diagnostic targets, phoB and pstA, for use in real-time PCR diagnostic assays. Real-time PCR diagnostic assays using these targets were designed and optimised to specifically detect and identify all 55H. influenzae strains tested. These novel rapid assays can be applied to the specific detection and identification of H. influenzae for use in epidemiological studies and could also enable improved monitoring of invasive disease caused by these bacteria.
流感嗜血杆菌被认为是一种重要的人类病原体，与包括血流感染和脑膜炎在内的侵袭性感染有关。目前使用的基于分子的诊断检测方法在正确检测和鉴定流感嗜血杆菌方面缺乏特异性。因此，需要开发用于特异性鉴定流感嗜血杆菌的新型诊断检测方法。进行了全基因组比较分析以鉴定推定的诊断靶点，这些靶点在流感嗜血杆菌的核苷酸序列中是独特的。通过该分析，我们鉴定出2个流感嗜血杆菌推定诊断靶点，即phoB和pstA，用于实时PCR诊断检测。使用这些靶点的实时PCR诊断检测方法被设计并优化，以特异性检测和鉴定所有测试的55株流感嗜血杆菌菌株。这些新型快速检测方法可应用于流感嗜血杆菌的特异性检测和鉴定，用于流行病学研究，还可改善对这些细菌引起的侵袭性疾病的监测。
