Seco Bruna Mara Silva, Campos Juliana Coutinho, da Costa Rocha Darlan Augusto, de Lima Aline Valerio, de Oliveira Fernanda Filomena, Lemo Mara Elisa Borsato, Sampaio Suely Carlos Ferreira, Sampaio Jorge Luiz Mello
School of Pharmacy, Clinical Microbiology and Antimicrobial Resistance Laboratory, University of São Paulo, Av. Professor Lineu Prestes, 580, São Paulo, SP, Zip code 05508-900, Brazil.
Fleury Medicine and Health, Microbiology Section, São Paulo, Brazil.
Braz J Microbiol. 2019 Jan;50(1):127-132. doi: 10.1007/s42770-018-0037-y. Epub 2018 Dec 6.
Carba-NP original report for blood cultures described the need of subculture and mechanical lysis before testing, reaching the turnaround time of approximately 4 hours for sample preparation. We tested 100 consecutive blood cultures positive for Gram-negative bacilli on the Gram stain from a large clinical laboratory. Bacterial pellets were prepared by centrifugation and submitted to Carba-NP and Blue-Carba tests and used further to prepare smears for Vitek MS. Results obtained with colonies grown on sheep blood agar using the same methodologies were used as the gold standard. Carbapenemase genes were confirmed by PCR and DNA sequencing. Vitek MS identified correctly 86% of the samples. Of note, 7% of the samples were incorrectly reported by the instrument as containing a single isolate. KPC-2 was the predominant carbapenemase detected. There was 100% concordance for both negative and positive results for Carba-NP. In contrast, for Blue-Carba the concordance for positive results was 92.8%, and 41% of strains negative for carbapenemases presented a yellowish color on control well turning the test non-interpretable. The turnaround time for sample preparation for preparing the pellet was 13 min, and no subculture or mechanical lysis is needed when detecting KPC production in Enterobacterales.
血培养的Carba-NP原始报告描述了在检测前需要进行传代培养和机械裂解,样本制备的周转时间约为4小时。我们对一家大型临床实验室革兰氏染色显示革兰氏阴性杆菌阳性的100份连续血培养样本进行了检测。通过离心制备细菌沉淀,并进行Carba-NP和Blue-Carba检测,进一步用于制备Vitek MS涂片。使用相同方法在羊血琼脂上生长的菌落所获得的结果用作金标准。通过PCR和DNA测序确认碳青霉烯酶基因。Vitek MS正确鉴定了86%的样本。值得注意的是,7%的样本被仪器错误报告为仅含有单一分离株。KPC-2是检测到的主要碳青霉烯酶。Carba-NP的阴性和阳性结果一致性均为100%。相比之下,对于Blue-Carba,阳性结果的一致性为92.8%,41%碳青霉烯酶阴性的菌株在对照孔呈现淡黄色,导致检测结果无法解读。制备沉淀的样本周转时间为13分钟,检测肠杆菌科细菌中KPC产生时无需传代培养或机械裂解。