Improved multiplex PCR primers for rapid identification of coagulase-negative staphylococci
- PDF / 848,608 Bytes
- 11 Pages / 595.276 x 790.866 pts Page_size
- 43 Downloads / 202 Views
ORIGINAL PAPER
Improved multiplex PCR primers for rapid identification of coagulase‑negative staphylococci Jonguk Kim1 · Jisoo Hong1 · Jeong‑A Lim1 · Sunggi Heu2 · Eunjung Roh1
Received: 7 July 2016 / Revised: 13 July 2017 / Accepted: 25 July 2017 © The Author(s) 2017. This article is an open access publication
Abstract Coagulase-negative staphylococci (CNS) are opportunistic pathogens that are currently emerging as causative agents of human disease. Though CNS are widespread in the clinic and food, their precise identification at species level is important. Here, using 16S rRNA sequencing, 55 staphylococcal isolates were identified as S. capitis, S. caprae, S. epidermidis, S. haemolyticus, S. pasteuri, S. saprophyticus, S. warneri, and S. xylosus. Although 16S rRNA sequencing is universally accepted as a standard for bacterial identification, the method did not effectively discriminate closely related species, and additional DNA sequencing was required. The divergence of the sodA gene sequence is higher than that of 16S rRNA. To devise a rapid and accurate identification method, sodA-specific primers were designed to demonstrate that species-specific multiplex polymerase chain reaction (PCR) can be used for the identification of CNS species. The accuracy of this method was higher than that of phenotypic identification; the method is simple and less time-consuming than 16S rRNA sequencing. Of the 55 CNS isolates, 92.72% were resistant to at least one antibiotic, and 60% were resistant to three or more Communicated by Jorge Membrillo-Hernández. Electronic supplementary material The online version of this article (doi:10.1007/s00203-017-1415-9) contains supplementary material, which is available to authorized users. * Eunjung Roh [email protected] 1
Microbial Safety Team, National Institute of Agricultural Sciences, Rural Development Administration, Wanju 55365, Republic of Korea
2
Crop Cultivation and Environment Research Division, National Institute of Crop Science, Rural Development Administration, Suwon 16613, Republic of Korea
antibiotics. CNS isolates produced diverse virulence-associated enzymes, including hemolysin (produced by 69.09% of the isolates), protease (65.45%), lipase (54.54%), lecithinase (36.36%), and DNase (29.09%); all isolates could form a biofilm. Because of the increasing pathogenic significance of CNS, the efficient multiplex PCR detection method developed in this study may contribute to studies for human health. Keywords Coagulase-negative staphylococci · Speciesspecific PCR · sodA · Antimicrobial resistance · Virulence factor
Introduction Staphylococci includes coagulase-positive staphylococci (CPS), almost exclusively represented by Staphylococcus aureus, and coagulase-negative staphylococci (CNS) (Becker et al. 2004). To date, the Staphylococcus genus comprises 49 species and 26 subspecies (Han et al. 2015). Most studies on staphylococcal pathogenicity have focused on S. aureus, and little attention has been paid to CNS (ChajeckaWierzchowska et al. 2015). CNS have been
Data Loading...
