Polymerase chain reaction-based screening for the ceftriaxone-resistant Neisseria gonorrhoeae F89 strain

Emergence and spread of Neisseria gonorrhoeae resistant to extended spectrum cephalosporins is a major problem threatening treatment of gonorrhoea and is further highlighted by the recent report of a second ceftriaxone-resistant N. gonorrhoeae strain (F89) in Europe, initially observed in France and...

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Published in:	Euro surveillance : bulletin européen sur les maladies transmissibles Vol. 18; no. 14; p. 20444
Main Authors:	Goire, N, Lahra, M M, Ohnishi, M, Hogan, T, Liminios, A E, Nissen, M D, Sloots, T P, Whiley, D M
Format:	Journal Article
Language:	English
Published:	Sweden Centre Europeen pour la Surveillance Epidemiologique du SIDA (European Centre for the Epidemiological Monitoring of AIDS) 04-04-2013
Subjects:	Anti-Bacterial Agents Antibiotics Ceftriaxone - pharmacology Drug resistance Drug Resistance, Bacterial Europe Gonorrhea Gram-negative bacteria Humans Microbial Sensitivity Tests Neisseria gonorrhoeae - drug effects Neisseria gonorrhoeae - genetics Neisseria gonorrhoeae - pathogenicity Penicillin-Binding Proteins - genetics Polymerase Chain Reaction Europe
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Summary:	Emergence and spread of Neisseria gonorrhoeae resistant to extended spectrum cephalosporins is a major problem threatening treatment of gonorrhoea and is further highlighted by the recent report of a second ceftriaxone-resistant N. gonorrhoeae strain (F89) in Europe, initially observed in France and subsequently identified in Spain. N. gonorrhoeae antimicrobial resistance (AMR) surveillance has acquired new importance and molecular tools have the potential to enhance bacterial culture-based methods. In this study, we established a polymerase chain reaction (PCR) protocol for direct detection of the F89 strain. A key component of this screening protocol was the development of a hybridisation probe-based melting curve analysis assay (mosaic501-hybPCR) to detect the presence of an A501P substitution on the N. gonorrhoeae mosaic penicillin binding protein 2 (PBP2) sequence, an important characteristic of the F89 strain. The mosaic501-hybPCR was evaluated using plasmid-derived positive controls (n=3) and characterised gonococcal (n=33) and non-gonococcal (n=58) isolates. The protocol was then applied to 159 clinical specimens from Sydney, Australia, collected during the first half of the year 2012 that were N. gonorrhoeae PCR-positive. Overall, the results indicate that the PCR-based protocol is suitable for direct detection of the N. gonorrhoeae F89 strain in non-cultured clinical samples. It therefore provides an additional tool to aid investigations into the potential spread of F89 strain throughout Europe and elsewhere.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1560-7917 1025-496X 1560-7917
DOI:	10.2807/1560-7917.es2013.18.14.20444