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Multiplex real-time...
Multiplex real-time PCR assay with high-resolution melting analysis for characterization of antimicrobial resistance in neisseria gonorrhoeae
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- Donà, Valentina (författare)
- Institute for Infectious Diseases, University of Bern, Bern, Switzerland
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- Kasraian, Sara (författare)
- Institute for Infectious Diseases, University of Bern, Bern, Switzerland
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- Lupo, Agnese (författare)
- Institute for Infectious Diseases, University of Bern, Bern, Switzerland
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- Guilarte, Yuvia N. (författare)
- Institute for Infectious Diseases, University of Bern, Bern, Switzerland
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- Hauser, Christoph (författare)
- Department of Infectious Diseases, Bern University Hospital, University of Bern, Bern, Switzerland
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- Furrer, Hansjakob (författare)
- Department of Infectious Diseases, Bern University Hospital, University of Bern, Bern, Switzerland
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- Unemo, Magnus, 1970- (författare)
- Örebro universitet,Institutionen för hälsovetenskaper
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- Low, Nicola (författare)
- Department of Infectious Diseases, Bern University Hospital, University of Bern, Bern, Switzerland
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- Endimiani, Andrea (författare)
- Institute for Infectious Diseases, University of Bern, Bern, Switzerlanda
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(creator_code:org_t)
- Washington, USA : American Society for Microbiology, 2016
- 2016
- Engelska.
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Ingår i: Journal of Clinical Microbiology. - Washington, USA : American Society for Microbiology. - 0095-1137 .- 1098-660X. ; 54:8, s. 2074-2081
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
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- Resistance to antibiotics used against Neisseria gonorrhoeae infections is a major public health concern. Antimicrobial resistance (AMR) testing relies on time-consuming culture-based methods. Development of rapid molecular tests for detecting AMR determinants could provide valuable tools for surveillance, epidemiological studies and to inform individual case management. We developed a fast (<1.5 hrs) SYBR-green based real-time PCR method with high resolution melting (HRM) analysis. One triplex and three duplex reactions included two sequences for N. gonorrhoeae identification and seven determinants of resistance to extended-spectrum cephalosporins (ESCs), azithromycin, ciprofloxacin, and spectinomycin. The method was validated by testing 39 previously fully-characterized N. gonorrhoeae strains, 19 commensal Neisseria spp., and an additional panel of 193 gonococcal isolates. Results were compared with culture-based AMR determination. The assay correctly identified N. gonorrhoeae and the presence or absence of the seven AMR determinants. There was some cross-reactivity with non-gonococcal Neisseria species and the detection limit was 10(3)-10(4) gDNA copies/reaction. Overall, the platform accurately detected resistance to ciprofloxacin (sensitivity and specificity, 100%), ceftriaxone (sensitivity 100%, specificity 90%), cefixime (sensitivity 92%, specificity 94%), azithromycin and spectinomycin (both sensitivity and specificity, 100%). In conclusion, our methodology accurately detects mutations generating resistance to antibiotics used to treat gonorrhea. Low assay sensitivity prevents direct diagnostic testing of clinical specimens but this method can be used to screen collections of gonococcal isolates for AMR more quickly than with current culture-based AMR testing.
Ämnesord
- NATURVETENSKAP -- Biologi -- Mikrobiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Microbiology (hsv//eng)
Nyckelord
- Microbiology
- Mikrobiologi
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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Till lärosätets databas
- Av författaren/redakt...
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Donà, Valentina
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Kasraian, Sara
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Lupo, Agnese
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Guilarte, Yuvia ...
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Hauser, Christop ...
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Furrer, Hansjako ...
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Unemo, Magnus, 1 ...
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Low, Nicola
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Endimiani, Andre ...
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- Om ämnet
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- NATURVETENSKAP
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NATURVETENSKAP
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och Biologi
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och Mikrobiologi
- Artiklar i publikationen
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Journal of Clini ...
- Av lärosätet
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Örebro universitet