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

被引：27

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

来源：

FUTURE MICROBIOLOGY | 2017年 / 12卷 / 07期

基金：

英国医学研究理事会; 澳大利亚国家健康与医学研究理事会;

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

页码：585 / 593

页数：9