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High-Resolution Genotyping of Chlamydia trachomatis by Use of a Novel Multilocus Typing DNA Microarray
Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinska vetenskaper, Klinisk bakteriologi.
Friedrich-Loeffler-Institut (Federal Research Institute for Animal Health), Institute of Molecular Pathogenesis, Jena, Germany.
Department of Microbiology and Infection Control, University Hospital of North Norway, Tromsø, Norway.
Alere Technologies GmbH, Jena, Germany.
Vise andre og tillknytning
2011 (engelsk)Inngår i: Journal of Clinical Microbiology, ISSN 0095-1137, E-ISSN 1098-660X, Vol. 49, nr 8, s. 2838-2843Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Typing of Chlamydia trachomatis is important to understandingits epidemiology. Currently used methods such as DNA sequencingof the ompA gene and multilocus sequence typing (MLST) eitheroffer limited epidemiological resolution or are laborious andexpensive, or both. DNA microarray technology using the ArrayStripformat is an affordable alternative for genotyping. In thisstudy, we developed a new multilocus typing (MLT) DNA microarray,based on the target regions of a high-resolution MLST systemas well as software for easy analysis. Validation of the arraywas done by typing 80 previously MLST-typed clinical specimensfrom unselected adolescents in school. The MLT array showed100% specificity and provided 2.4-times-higher resolution thanompA sequencing, separating the commonly predominating ompAE/Bour genotype into 7 MLT array genotypes. The MLT array reproducedepidemiological findings revealed by the MLST system and showedsufficient sensitivity to work with clinical specimens. Comparedto MLST analysis, the expenses needed for testing a sample withthe MLT array are considerably lower. Moreover, testing canbe completed within 1 working day rather than 3 or 4 days, withdata analysis not requiring highly specialized personnel. Thepresent MLT array represents a powerful alternative in C. trachomatisgenotyping.

sted, utgiver, år, opplag, sider
2011. Vol. 49, nr 8, s. 2838-2843
Emneord [en]
Chlamydia trachomatis, genotyping, microarray, MLT array, multilocus sequence typing (MLST), multilocus typing array, ompA
HSV kategori
Identifikatorer
URN: urn:nbn:se:uu:diva-156746DOI: 10.1128/JCM.00883-11ISI: 000293221900009OAI: oai:DiVA.org:uu-156746DiVA, id: diva2:433138
Tilgjengelig fra: 2011-08-09 Laget: 2011-08-09 Sist oppdatert: 2017-12-08bibliografisk kontrollert
Inngår i avhandling
1. High Resolution Genotyping of Chlamydia trachomatis
Åpne denne publikasjonen i ny fane eller vindu >>High Resolution Genotyping of Chlamydia trachomatis
2011 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Chlamydia trachomatis is an obligate intracellular bacterium of major human health concern, causing urogential chlamydia infections, lymphogranuloma venereum (LGV) and trachoma. Chlamydia is one of the most common sexually transmitted infections worldwide and can cause infertility.

In the first four papers described herein we used a high resolution multilocus sequence typing (MLST) system to investigate the epidemiology of C. trachomatis, and showed that MLST is superior to conventional ompA genotyping with respect to resolution. In the fifth paper we simplified the methodology by developing and validating a multilocus typing (MLT) DNA microarray based on the MLST system.

In more detail, MLST analysis of consecutive specimens from 2006 in Örebro County in Sweden, and comparison to specimens from 1999-2000, showed that the new variant C. trachomatis (nvCT) is monoclonal and likely has appeared in recent years.

MLST analysis of LGV specimens from men who have sex with men (MSM) showed that the increase of LGV in Europe in the last decade indeed was a clonal outbreak, contrary to the USA where LGV might have been present all along.

In the third paper, clinical symptoms could not be correlated with the MLST genotypes, suggesting, together with the combined results of all previous studies, that bacterial factors, if important, need to be understood in the context of host factors.

MLST analysis of specimens from a high incidence C. trachomatis area in North Norway revealed interesting epidemiological details concerning unusual genetic variants, the nvCT and MSM, but found no significant difference in genetic diversity compared to two other geographic areas in Norway.

Lastly, we developed a MLT array that provides high resolution while being rapid and cost-effective, which makes it an interesting alternative for C. trachomatis genotyping.

In conclusion, the MLST system and the MLT array have proven to be useful tools and should now be applied in further investigations to improve our understanding of C. trachomatis epidemiology.

sted, utgiver, år, opplag, sider
Uppsala: Acta Universitatis Upsaliensis, 2011. s. 48
Serie
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 690
Emneord
Chlamydia trachomatis: multilocus sequence typing: MLST: genotyping: lymphogranuloma venereum: new variant C. tracomatis: nvCT: multilocus typing DNA microarray: MLT array
HSV kategori
Forskningsprogram
Medicinsk vetenskap
Identifikatorer
urn:nbn:se:uu:diva-156751 (URN)978-91-554-8121-6 (ISBN)
Disputas
2011-09-29, Hörsalen, Klinisk mikrobiologi, Akademiska sjukhuset, Dag Hammarskjöldsväg 17, Uppsala, 09:15 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2011-08-30 Laget: 2011-08-09 Sist oppdatert: 2018-01-12

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