Laboratory for Mass Spectrometry and Functional Proteomics, Rudjer Bošković Institute, Zagreb, Croatia.
Laboratory for Aquaculture Biotechnology, Rudjer Bošković Institute, Zagreb, Croatia.
J Fish Dis. 2019 Aug;42(8):1201-1209. doi: 10.1111/jfd.13022. Epub 2019 Jun 13.
MALDI-TOF MS was tested for the identification of Photobacterium damselae subsp. piscicida on isolates grown on two media, cultured at three incubation times and applied on the target plate by the direct sample spotting (DS), by the on-target extraction (OTE) and by the full extraction (FE) method, in triplicates. The identification of samples grown on blood agar (BA) outperformed identification on tryptic soya agar (TSA) by 0.64% for DS and OTE. The OTE gave the highest scores in both culture media, all incubation times and replicates. Reliable 24-hr species identification was 61.54%, 84.61% and 53.85% for samples grown on TSA and identified by DS, OTE and FE, respectively. For isolates grown on BA, they were 76.92%, 96.15% and 30.77%, respectively. When identified by OTE, the 48-hr identification was 93.58%, but for 72 hr declined to 71.79%. The reliable identification with the highest score from the first measurement was 100% only for OTE from BA (24 hr), whereas OTE from TSA gave 84.61% (24 hr), 76.92% (48 hr) and 84.61% (72 hr). The reliable MALDI-TOF MS identification of Ph. damselae subsp. piscicida is incubation time, media, target plate preparation and replicate-dependent.
基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)用于鉴定两种培养基上生长的 Photobacterium damselae subsp. piscicida 分离株,培养三个时间点,采用直接样本点样(DS)、目标萃取(OTE)和全萃取(FE)三种方法将培养物加在目标板上,每个样本重复三次。DS 和 OTE 鉴定血琼脂(BA)上生长的样本比鉴定在胰蛋白酶大豆琼脂(TSA)上的鉴定结果分别高出 0.64%。OTE 在两种培养基、所有培养时间和重复中均给出了最高的评分。在 TSA 上生长并通过 DS、OTE 和 FE 鉴定的样本,可靠的 24 小时种属鉴定率分别为 61.54%、84.61%和 53.85%。在 BA 上生长的分离株,其分别为 76.92%、96.15%和 30.77%。OTE 鉴定的 48 小时鉴定率为 93.58%,但 72 小时后下降至 71.79%。只有 OTE 从 BA(24 小时)得到的第一次测量的最高评分的可靠鉴定率为 100%,而 OTE 从 TSA 得到的可靠鉴定率分别为 84.61%(24 小时)、76.92%(48 小时)和 84.61%(72 小时)。Ph. damselae subsp. piscicida 的可靠 MALDI-TOF MS 鉴定依赖于培养时间、培养基、目标板准备和重复。