Institute of Medical Microbiology, University Hospital Münster, Münster, Germany.
Friedrich Loeffler-Institute of Medical Microbiology, University Medicine Greifswald, Greifswald, Germany.
Microbiol Spectr. 2021 Sep 3;9(1):e0003821. doi: 10.1128/Spectrum.00038-21. Epub 2021 Jun 9.
Short incubation of positive blood cultures on solid media is now increasingly applied to speed up species identification by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Although Columbia blood agar (CBA) and chocolate agar (Choc) are widely used, a direct comparison of standard agars is lacking. We therefore compared the time to species identification of blood cultures incubated on CBA, Choc, and MacConkey agar (MAC, for Gram-negative rods). Positive aerobic/anaerobic blood cultures (2 drops = 50 μl) were incubated on CBA, Choc, MAC, and the required time of incubation to low-confidence identification (score of ≥1.7 to <2) and high-confidence identification (score of ≥2) by MALDI-TOF MS was measured. Exclusion criteria were (i) false-positive blood cultures, (ii) mixed cultures with different species, (iii) growth of anaerobes/fungi, and (iv) a total number of isolates of one group (i.e., Gram-positive/-negative cocci/rods) of <30. A total of 187 blood cultures with Gram-positive cocci ( = 124) and Gram-negative rods ( = 63) were included in the final analysis. The shortest median time to high-confidence identification (score of ≥2) was achieved on MAC for Gram-negative rods (2.0 h; range, 1.9 to 4.2 h) and on CBA for Gram-positive cocci (4.0 h; range, 1.9 to 25.0 h). However, the difference from results obtained with Choc was not statistically significant. When only one agar plate is used for short incubation of positive blood cultures, Choc may represent a compromise in terms of time to high-confidence identification by MALDI-TOF MS and the bacterial spectrum that is covered. However, using only Choc is disadvantageous when the shortest incubation times to identification are strived for. When blood cultures are flagged as positive, they are incubated on solid media to produce enough biomass of the bacterium for identification and susceptibility testing. Rapid turnaround times for laboratory results could save lives, and we wanted to assess which solid medium is best to shorten the time to species identification using MALDI-TOF mass spectrometry. For that purpose, we used positive blood cultures from routine diagnostics and compared Columbia blood agar (CBA), Chocolate agar (Choc), and MacConkey agar (MAC, for Gram-negative rods). We found that MAC performed best for Gram-negative rods and CBA was quickest for Gram-positive cocci. However, Choc may represent a compromise if fastidious species should be covered.
短时间孵育阳性血培养物在固体培养基上现在越来越多地被用于通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)来加速物种鉴定。虽然哥伦比亚血琼脂(CBA)和巧克力琼脂(Choc)被广泛使用,但缺乏对标准琼脂的直接比较。因此,我们比较了在 CBA、Choc 和麦康凯琼脂(MAC,用于革兰氏阴性杆菌)上孵育的血培养物达到种属鉴定的时间。将有氧/厌氧血培养物(2 滴 = 50 μl)接种于 CBA、Choc、MAC,并测量通过 MALDI-TOF MS 获得低置信度鉴定(评分≥1.7 至<2)和高置信度鉴定(评分≥2)所需的时间。排除标准为:(i)假阳性血培养物,(ii)不同物种的混合培养物,(iii)厌氧菌/真菌的生长,以及(iv)一组(即革兰氏阳性/阴性球菌/杆菌)的分离物总数<30。最终分析共纳入 187 株革兰氏阳性球菌( = 124)和革兰氏阴性杆菌( = 63)血培养物。获得高置信度鉴定(评分≥2)的最短中位时间是在 MAC 上用于革兰氏阴性杆菌(2.0 h;范围,1.9 至 4.2 h)和在 CBA 上用于革兰氏阳性球菌(4.0 h;范围,1.9 至 25.0 h)。然而,与 Choc 的结果差异没有统计学意义。当仅使用一个琼脂平板进行阳性血培养物的短时间孵育时,Choc 可能代表通过 MALDI-TOF MS 进行高置信度鉴定和涵盖的细菌谱的折衷。然而,当力求达到最短的鉴定孵育时间时,仅使用 Choc 是不利的。当血液培养物被标记为阳性时,它们被接种在固体培养基上以产生足够的细菌生物量进行鉴定和药敏试验。实验室结果的快速周转时间可以挽救生命,我们想评估哪种固体培养基最适合使用 MALDI-TOF 质谱缩短种属鉴定时间。为此,我们使用了常规诊断中的阳性血培养物,并比较了哥伦比亚血琼脂(CBA)、巧克力琼脂(Choc)和麦康凯琼脂(MAC,用于革兰氏阴性杆菌)。我们发现 MAC 对革兰氏阴性杆菌的效果最好,而 CBA 对革兰氏阳性球菌的鉴定速度最快。然而,如果需要涵盖苛刻的物种,Choc 可能是一个妥协。