通过基质辅助激光解吸电离飞行时间质谱分析实现产β-内酰胺酶肠杆菌科细菌的早期检测。
Towards the early detection of β-lactamase-producing Enterobacteriaceae by MALDI-TOF MS analysis.
作者信息
Oviaño Marina, Gómara Marta, Barba Maria José, Revillo María José, Barbeyto Luis Pedro, Bou Germán
机构信息
Servicio de Microbiología, Complejo Hospitalario Universitario A Coruña, La Coruña, Spain.
Servicio de Microbiología, Hospital Universitario Miguel Servet, Zaragoza, Spain.
出版信息
J Antimicrob Chemother. 2017 Aug 1;72(8):2259-2262. doi: 10.1093/jac/dkx127.
OBJECTIVES
Development of a novel MALDI-TOF MS-based method for the rapid detection of ESBL-producing Enterobacteriaceae.
METHODS
The method was evaluated in terms of sensitivity, specificity and turnaround time regarding the antibiotic used (cefotaxime, ceftazidime, ceftriaxone, cefpodoxime or cefepime) and the performance of the automated MBT STAR-BL software (Bruker Daltonik GmbH, Germany) relative to qualitative interpretation of spectra for detecting β-lactamase resistance by MALDI-TOF MS (Bruker Daltonik) in a collection of 11 isogenic Escherichia coli control strains expressing different β-lactamases. Finally, for clinical validation, β-lactamase activity was determined under previously evaluated conditions in 100 clinical isolates previously characterized by PCR and sequencing.
RESULTS
Clinical validation of the assay showed 100% sensitivity and specificity for detecting β-lactam resistance in 30 min by measuring hydrolysis of ceftriaxone (0.50 mg/mL) with the automated MBT STAR-BL software. Regarding the antibiotics evaluated, ceftriaxone yielded 70% more positive results than cefotaxime, 80% more than ceftazidime and 20% more than cefpodoxime, with 100% specificity. Cefepime revealed 100% sensitivity, but only 27% specificity. For the same incubation time, the automated software yielded on average 41% more positive results in relation to detecting resistance than qualitative interpretation of spectra.
CONCLUSIONS
Our clinical validation of the method proved it to be highly reliable, simple to perform and time saving, transforming β-lactam resistance detection by MALDI-TOF MS into a ready-to-use technique in clinical laboratories.
目的
开发一种基于基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）的新方法，用于快速检测产超广谱β-内酰胺酶（ESBL）的肠杆菌科细菌。
方法
针对所使用的抗生素（头孢噻肟、头孢他啶、头孢曲松、头孢泊肟或头孢吡肟），以及在11株表达不同β-内酰胺酶的同基因大肠杆菌对照菌株中，通过MALDI-TOF MS（布鲁克道尔顿公司）检测β-内酰胺酶耐药性时，自动MBT STAR-BL软件（德国布鲁克道尔顿公司）相对于光谱定性解读的性能，对该方法的灵敏度、特异性和周转时间进行评估。最后，为进行临床验证，在先前评估的条件下，对100株先前通过聚合酶链反应（PCR）和测序进行鉴定的临床分离株测定β-内酰胺酶活性。
结果
该检测方法的临床验证表明，使用自动MBT STAR-BL软件通过测量头孢曲松（0.50 mg/mL）的水解情况，在30分钟内检测β-内酰胺耐药性的灵敏度和特异性均为100%。在所评估的抗生素中，头孢曲松产生的阳性结果比头孢噻肟多70%，比头孢他啶多80%，比头孢泊肟多20%，特异性为100%。头孢吡肟显示出100%的灵敏度，但特异性仅为27%。在相同的孵育时间内，与光谱定性解读相比，自动软件检测耐药性时产生的阳性结果平均多41%。
结论
我们对该方法的临床验证证明其高度可靠、操作简单且节省时间，将通过MALDI-TOF MS检测β-内酰胺耐药性转变为临床实验室中一种易于使用的技术。
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