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目的比较基质辅助激光解吸/电离飞行时间质谱(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF MS)直接涂片法和蛋白提取法鉴定洋葱伯克霍尔德菌群的差异。方法收集118株临床分离经VITEK全自动细菌仪鉴定为洋葱伯克霍尔德菌群菌株,采用MALDI-TOF MS直接涂片法和蛋白提取法进一步分析基因型,同时比较两种鉴定方法之间的优缺点。结果 118株洋葱伯克霍尔德菌群经MALDI-TOF MS两种方法只是鉴定结果分数有所差异,鉴定结果一致,分别为:93株Burkhoderia cencepacia(基因型Ⅲ型),11株多嗜伯克霍尔德菌(基因型Ⅱ型),10株洋葱伯克霍尔德菌(基因型I型),3株越南伯克霍尔德菌(基因型V型),1株吡咯伯克霍尔德菌(基因型Ⅸ型)。MALDI-TOF MS直接涂片法鉴定分数均值2.126,仅9株(7.62%)鉴定结果不可信(分值<1.700),准确鉴定至种(t≥2.300)与准确鉴定至属(t≥1.700)的比例分别为38.98%和92.38%;MALDI-TOF MS蛋白提取法仅2株(1.69%)鉴定结果不可信,准确鉴定至种(t≥2.300)与准确鉴定至属(t≥1.700)的百分比均高于直接涂片法,分别为44.92%和98.31%;但是不同分值分组间准确鉴定至种(分值≥2.300)、鉴定至属(分值≥1.700),两种方法差异均无统计学意义(P>0.05)。结论 MALDI-TOF MS可快速准确地鉴定洋葱伯克霍尔德菌群至基因型,临床标本鉴定时首选MALDI-TOF MS直接涂片法,对于鉴定结果不可信的菌株可进一步采用蛋白提取法鉴定。
OBJECTIVE To compare the expression of Burkholderia cepacia by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS) direct smear and protein extraction difference. Methods 118 clinical isolates were identified as Burkholderia cepacia by VITEK automatic bacterial analyzer. The genotypes were further analyzed by MALDI-TOF MS direct smear method and protein extraction method, and the differences between the two methods were compared. The advantages and disadvantages. Results The 118 Burkholderia cepacia populations were identified by MALDI-TOF MS. The results were consistent with those of Burkholderia cepacia (93 Bur Burkoderia cencepacia, 11 genotypes) Klebsiella (genotype II), 10 Burkholderia cepacia (genotype I), 3 Burkholderia cepacia (genotype V), and 1 ppy Burkhol Bacteria (genotype Ⅸ). MALDI-TOF MS direct smear method identified a mean score of 2.126, only 9 strains (7.62%) were unreliable (score <1.700), accurately identified to species (t≥2.300) and accurately identified to belong to (t≥1.700) (38.98%) and 92.38% (92.38%) respectively. Only 2 strains (1.69%) of MALDI-TOF MS protein extraction method were unreliable and the percentages of the species (t≥2.300) Were higher than the direct smear method, respectively, 44.92% and 98.31%; but different scores between groups accurately identified to species (score ≥ 2.300), identified to the genus (score ≥ 1.700), no difference between the two methods Significance (P> 0.05). Conclusion MALDI-TOF MS can rapidly and accurately identify Burkholderia cepacia to genotype. MALDI-TOF MS direct smear is the first choice for clinical specimen identification. The unidentified strains can be further identified by protein extraction .