Evaluation of performance of the GENECUBE assay for rapid molecular identification of Staphylococcus aureus and methicillin resistance in positive blood culture medium
Evaluation of performance of the GENECUBE assay for rapid molecular identification of Staphylococcus aureus and methicillin resistance in positive blood culture medium
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DOI：
10.1371/journal.pone.0219819
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发表时间：
2019-07-16
期刊：
影响因子：
3.7
通讯作者：
Suzuki, Hiromichi
中科院分区：
文献类型：
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作者：
Hida, Yukio;Uemura, Keiichi;Suzuki, Hiromichi
从阳性血培养介质中快速鉴定病原体是及时靶向治疗脓毒症患者的前提。The GENECUBE(TOYOBO Co.，Ltd.)是一种新型的全自动基因分析仪，可以提纯DNA并扩增目标DNA。在这项研究中，我们评估了两种新发展的GENECUBE方法直接检测血培养上清液中nuc和mecA基因的能力；nuc是金黄色葡萄球菌所特有的，mecA表明对甲氧西林耐药。我们检查了从三家医院采集的263份阳性血培养样本，这些样本来自怀疑患有葡萄球菌菌血症的患者。然后将结果与基质辅助激光解吸/电离飞行时间质谱仪、药物敏感性测试(MicroScan系统或干板Eiken)和测序分析结果进行比较。GENECUBE法检测血培养阳性金黄色葡萄球菌和甲氧西林耐药性的敏感性和特异性均为100%。对36份血培养阳性样本的检查周转时间进行了评估。从开始孵育到完成GENECUBE检查的时间为23h(四分位数范围：IQR 21-37h)，报告革兰氏染色检查到完成GENECUBE检查的时间为52min(IQR 48-62min)。这些发现表明，GENECUBE检测显著缩短了检测时间，而不损失敏感性或特异性。
Rapid identification of causative agents from positive blood culture media is a prerequisite for the timely targeted treatment of patients with sepsis. The GENECUBE (TOYOBO Co., Ltd.) is a novel, fully-automated gene analyzer that can purify DNAs and amplify target DNAs. In this study, we evaluated the ability of two newly developed GENECUBE assays to directly detect the nuc and mecA genes in blood culture medium; nuc is specific to Staphylococcus aureus, and mecA indicates methicillin resistance. We examined 263 positive blood culture samples taken at three hospitals from patients suspected of having staphylococcal bacteremia. The results were then compared with those obtained using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, antimicrobial susceptibility testing (Microscan system or Dry-plate EIKEN), and sequencing analysis. The GENECUBE assays had sensitivity and specificity of 100% in detecting both S. aureus and methicillin resistance in positive blood culture. The turnaround time of the examination was evaluated for 36 positive blood culture samples. The time between the initiation of incubation and completion of the GENECUBE examination was 23 h (inter-quartile range: IQR 21-37 h); the time between reporting of Gram stain examination and completion of the GENECUBE examination was 52 min (IQR 48-62 min). These findings show that the GENECUBE assays significantly reduce the assay time with no loss of sensitivity or specificity.