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. 2014 Apr 4;9(4):e94064.
doi: 10.1371/journal.pone.0094064. eCollection 2014.

Evaluation of an automated rapid diagnostic assay for detection of Gram-negative bacteria and their drug-resistance genes in positive blood cultures

Masayoshi Tojo  1 , Takahiro Fujita  2 , Yusuke Ainoda  2 , Maki Nagamatsu  1 , Kayoko Hayakawa  3 , Kazuhisa Mezaki  4 , Aki Sakurai  5 , Yoshinori Masui  6 , Hirohisa Yazaki  7 , Hiroshi Takahashi  8 , Tohru Miyoshi-Akiyama  9 , Kyoichi Totsuka  2 , Teruo Kirikae  9 , Norio Ohmagari  3
Affiliations

Evaluation of an automated rapid diagnostic assay for detection of Gram-negative bacteria and their drug-resistance genes in positive blood cultures

Masayoshi Tojo et al. PLoS One. .

Abstract

We evaluated the performance of the Verigene Gram-Negative Blood Culture Nucleic Acid Test (BC-GN; Nanosphere, Northbrook, IL, USA), an automated multiplex assay for rapid identification of positive blood cultures caused by 9 Gram-negative bacteria (GNB) and for detection of 9 genes associated with β-lactam resistance. The BC-GN assay can be performed directly from positive blood cultures with 5 minutes of hands-on and 2 hours of run time per sample. A total of 397 GNB positive blood cultures were analyzed using the BC-GN assay. Of the 397 samples, 295 were simulated samples prepared by inoculating GNB into blood culture bottles, and the remaining were clinical samples from 102 patients with positive blood cultures. Aliquots of the positive blood cultures were tested by the BC-GN assay. The results of bacterial identification between the BC-GN assay and standard laboratory methods were as follows: Acinetobacter spp. (39 isolates for the BC-GN assay/39 for the standard methods), Citrobacter spp. (7/7), Escherichia coli (87/87), Klebsiella oxytoca (13/13), and Proteus spp. (11/11); Enterobacter spp. (29/30); Klebsiella pneumoniae (62/72); Pseudomonas aeruginosa (124/125); and Serratia marcescens (18/21); respectively. From the 102 clinical samples, 104 bacterial species were identified with the BC-GN assay, whereas 110 were identified with the standard methods. The BC-GN assay also detected all β-lactam resistance genes tested (233 genes), including 54 bla(CTX-M), 119 bla(IMP), 8 bla(KPC), 16 bla(NDM), 24 bla(OXA-23), 1 bla(OXA-24/40), 1 bla(OXA-48), 4 bla(OXA-58), and 6 blaVIM. The data shows that the BC-GN assay provides rapid detection of GNB and β-lactam resistance genes in positive blood cultures and has the potential to contributing to optimal patient management by earlier detection of major antimicrobial resistance genes.

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Conflict of interest statement

Competing Interests: H. Takahashi is a managing partner of East-West Diagnostics/Theranostics, LLC, and a consultant to Nanosphere. H. Takahashi has no other declarations relating to patents, products in development or marketed products. This study is not funded by these companies. The remaining authors have no reported potential conflicts of interest. This does not alter the authors’ adherence to all the PLOS ONE policies on sharing data and materials.

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