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Whole-genome sequences of Chlamydia trachomatis directly from clinical samples without culture

Seth-Smith, Helena M. B. (författare)
Wellcome Trust Sanger Inst, Hinxton, England
Harris, Simon R. (författare)
Wellcome Trust Sanger Inst, Hinxton, England
Skilton, Rachel J. (författare)
Fac Med, Mol Microbiol Grp, Southampton Gen Hosp, Univ Southampton, Southampton, England
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Radebe, Frans M. (författare)
Natl Inst Communicable Dis, Ctr HIV & Sexually Transmitted Infect, National Health Laboratory Service, Johannesburg, South Africa
Golparian, Daniel, 1984- (författare)
Natl Reference Lab Pathogen Neisseria, WHO Collaborating Ctr Gonorrhoea & Other STIs, Örebro University Hospital, Örebro, Sweden
Shipitsyna, Elena (författare)
Lab Microbiol, DO Ott Res Inst Obstet & Gynaecol, St Petersburg, Russia
Duy, Pham Thanh (författare)
Scott, Paul (författare)
Wellcome Trust Sanger Inst, Hinxton, England
Cutcliffe, Lesley T. (författare)
Fac Med, Mol Microbiol Grp, Southampton Gen Hosp, Univ Southampton, Southampton, England
O'Neill, Colette (författare)
Fac Med, Mol Microbiol Grp, Southampton Gen Hosp, Univ Southampton, Southampton, England
Parmar, Surendra (författare)
Hlth Protect Agcy, Clin Microbiol & Publ Hlth Lab, Addenbrookes Hosp, Cambridge, England
Pitt, Rachel (författare)
Sexually Transmitted Bacteria Reference Lab, Health Protection Agency, London, England
Baker, Stephen (författare)
Clin Res Unit, Wellcome Trust Major Overseas List, Hosp Trop Dis, Univ Oxford, Ho Chi Minh City, Vietnam
Ison, Catherine A. (författare)
Sexually Transmitted Bacteria Reference Lab, Health Protection AgencyLondon NW9 5HT, England.
Marsh, Peter (författare)
Publ Hlth Lab Southampton, Health Protection Agency, Southampton Gen Hosp, Southampton, England
Jalal, Hamid (författare)
Health Protection Agency, Clin Microbiol & Publ Hlth Lab, Addenbrookes Hosp, Cambridge, England
Lewis, David A. (författare)
Natl Inst Communicable Dis, Ctr HIV & Sexually Transmitted Infect, National Health Laboratory ServiceJohannesburg, South Africa; Fac Hlth Sci, Dept Internal Med, Univ Witwatersrand, Johannesburg, South Africa
Unemo, Magnus (författare)
Region Örebro län,Natl Reference Lab Pathogen Neisseria, WHO Collaborating Ctr Gonorrhoea & Other STIs, Örebro University Hospital, Örebro, Sweden
Clarke, Ian N. (författare)
Fac Med, Mol Microbiol Grp, Southampton Gen Hosp, Univ Southampton, Southampton, England
Parkhill, Julian (författare)
Wellcome Trust Sanger Inst, Hinxton, England
Thomson, Nicholas R. (författare)
Wellcome Trust Sanger Inst, Hinxton, England
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 (creator_code:org_t)
2013-03-22
2013
Engelska.
Ingår i: Genome Research. - : Cold Spring Harbor Laboratory Press (CSHL). - 1088-9051 .- 1549-5469. ; 23:5, s. 855-866
Relaterad länk:
http://genome.cshlp....
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https://urn.kb.se/re...
https://doi.org/10.1...
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  • Tidskriftsartikel (refereegranskat)
Abstract Ämnesord
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  • The use of whole-genome sequencing as a tool for the study of infectious bacteria is of growing clinical interest. Chlamydia trachomatis is responsible for sexually transmitted infections and the blinding disease trachoma, which affect hundreds of millions of people worldwide. Recombination is widespread within the genome of C. trachomatis, thus whole-genome sequencing is necessary to understand the evolution, diversity, and epidemiology of this pathogen. Culture of C trachomatis has, until now, been a prerequisite to obtain DNA for whole-genome sequencing; however, as C trachomatis is an obligate intracellular pathogen, this procedure is technically demanding and time consuming. Discarded clinical samples represent a large resource for sequencing the genomes of pathogens, yet clinical swabs frequently contain very low levels of C trachomatis DNA and large amounts of contaminating microbial and human DNA. To determine whether it is possible to obtain whole-genome sequences from bacteria without the need for culture, we have devised an approach that combines immunomagnetic separation (IMS) for targeted bacterial enrichment with multiple displacement amplification (MDA) for whole-genome amplification. Using IMS-MDA ill conjunction with high-throughput multiplexed Illumina sequencing, we have produced the first whole bacterial genome sequences direct from clinical samples. We also show that this method can be used to generate genome data from nonviable archived samples. This method will prove a useful tool in answering questions relating to the biology of many difficult-to-culture or fastidious bacteria of clinical concern.

Ämnesord

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
MEDICAL AND HEALTH SCIENCES  -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)

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