Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR

被引:28
作者
Price, Erin P. [1 ,2 ]
Harris, Tegan M. [1 ]
Spargo, Jessie C. [1 ]
Nosworthy, Elizabeth [1 ]
Beissbarth, Jemima [1 ]
Chang, Anne B. [1 ]
Smith-Vaughan, Heidi C. [1 ]
Sarovich, Derek S. [1 ,2 ]
机构
[1] Menzies Sch Hlth Res, Child Hlth Div, Darwin, NT, Australia
[2] Univ Sunshine Coast, Fac Sci Hlth Educ & Engn, Ctr Anim Hlth Innovat, Sippy Downs, Qld, Australia
基金
澳大利亚国家健康与医学研究理事会; 英国医学研究理事会;
关键词
fucP; genotyping; Haemophilus haemolyticus; Haemophilus influenzae; hypD; multiplex; real-time PCR; siaT; TaqMan; triplex; COMPARATIVE GENOMICS; COMPLETE DELETION; SEQUENCE; VIRULENCE; CARRIAGE; PATHOGEN; STRAINS; DISEASE; VACCINE;
D O I
10.2217/fmb-2016-0215
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Aim: To design a highly specific and sensitive multiplex real-time PCR assay for the differentiation of the pathogen Haemophilus influenzae from its nonpathogenic nearneighbor Haemophilus haemolyticus. Materials & methods: A comparison of 380 Haemophilus spp. genomes was used to identify loci specific for each species. Novel PCR assays targeting H. haemolyticus (hypD) and H. influenzae (siaT) were designed. Results & discussion: PCR screening across 143 isolates demonstrated 100% specificity for hypD and siaT. These two assays were multiplexed with the recently described fucP assay for further differentiation among H. influenzae. Conclusion: The triplex assay provides rapid, unambiguous, sensitive and highly specific genotyping results for the simultaneous detection of hypD and siaT, including fucose-positive H. influenzae (fucP), in a single PCR.
引用
收藏
页码:585 / 593
页数:9
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