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

来源：

FUTURE MICROBIOLOGY | 2017年 / 12卷 / 07期

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

页码：585 / 593

页数：9

共 42 条

[1] Minor groove binder-conjugated DNA probes for quantitative DNA detection by hybridization-triggered fluorescence [J].

Afonina, IA ;

Reed, MW ;

Lusby, E ;

Shishkina, IG ;

Belousov, YS .

BIOTECHNIQUES, 2002, 32 (04) :940-+

[2] Haemophilus haemolyticus Isolates Causing Clinical Disease [J].

Anderson, Raydel ;

Wang, Xin ;

Briere, Elizabeth C. ;

Katz, Lee S. ;

Cohn, Amanda C. ;

Clark, Thomas A. ;

Messonnier, Nancy E. ;

Mayer, Leonard W. .

JOURNAL OF CLINICAL MICROBIOLOGY, 2012, 50 (07) :2462-2465

[3] A biphasic epigenetic switch controls immunoevasion, virulence and niche adaptation in non-typeable Haemophilus influenzae [J].

Atack, John M. ;

Srikhanta, Yogitha N. ;

Fox, Kate L. ;

Jurcisek, Joseph A. ;

Brockman, Kenneth L. ;

Clark, Tyson A. ;

Boitano, Matthew ;

Power, Peter M. ;

Jen, Freda E-C. ;

McEwan, Alastair G. ;

Grimmond, Sean M. ;

Smith, Arnold L. ;

Barenkamp, Stephen J. ;

Korlach, Jonas ;

Bakaletz, Lauren O. ;

Jennings, Michael P. .

NATURE COMMUNICATIONS, 2015, 6

[4] Molecular Surveillance of True Nontypeable Haemophilus influenzae: An Evaluation of PCR Screening Assays [J].

Binks, Michael J. ;

Temple, Beth ;

Kirkham, Lea-Ann ;

Wiertsema, Selma P. ;

Dunne, Eileen M. ;

Richmond, Peter C. ;

Marsh, Robyn L. ;

Leach, Amanda J. ;

Smith-Vaughan, Heidi C. .

PLOS ONE, 2012, 7 (03)

[5] IMMUNIZATION OF UNITED-STATES CHILDREN WITH HEMOPHILUS-INFLUENZAE TYPE-B POLYSACCHARIDE VACCINE - A COST-EFFECTIVENESS MODEL OF STRATEGY ASSESSMENT [J].

COCHI, SL ;

BROOME, CV ;

HIGHTOWER, AW .

JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION, 1985, 253 (04) :521-529

[6] Structure of a fucose transporter in an outward-open conformation [J].

Dang, Shangyu ;

Sun, Linfeng ;

Huang, Yongjian ;

Lu, Feiran ;

Liu, Yufeng ;

Gong, Haipeng ;

Wang, Jiawei ;

Yan, Nieng .

NATURE, 2010, 467 (7316) :734-U130

[7] Genome sequencing of disease and carriage isolates of nontypeable Haemophilus influenzae identifies discrete population structure [J].

De Chiara, Matteo ;

Hood, Derek ;

Muzzi, Alessandro ;

Pickard, Derek J. ;

Perkins, Tim ;

Pizza, Mariagrazia ;

Dougan, Gordon ;

Rappuoli, Rino ;

Moxon, E. Richard ;

Soriani, Marco ;

Donati, Claudio .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2014, 111 (14) :5439-5444

[8] Complete Deletion of the Fucose Operon in Haemophilus influenzae Is Associated with a Cluster in Multilocus Sequence Analysis-Based Phylogenetic Group II Related to Haemophilus haemolyticus: Implications for Identification and Typing [J].

de Gier, Camilla ;

Kirkham, Lea-Ann S. ;

Norskov-Lauritsen, Niels .

JOURNAL OF CLINICAL MICROBIOLOGY, 2015, 53 (12) :3773-3778

[9] Characterization of Nontypable Haemophilus influenzae Isolates Recovered from Adult Patients with Underlying Chronic Lung Disease Reveals Genotypic and Phenotypic Traits Associated with Persistent Infection [J].

Garmendia, Junkal ;

Viadas, Cristina ;

Calatayud, Laura ;

Mell, Joshua Chang ;

Marti-Lliteras, Pau ;

Euba, Begona ;

Llobet, Enrique ;

Gil, Carmen ;

Antonio Bengoechea, Jose ;

Redfield, Rosemary J. ;

Linares, Josefina .

PLOS ONE, 2014, 9 (05)

[10]

Giufre M, 2015, GENOME ANNOUNC, V3

← 1 2 3 4 5 →