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目的报道1例我国河北地区由格特隐球菌引起的脑膜炎,并对该菌进行试验研究。方法将分离自患者脑脊液中的隐球菌分别接种于Chromagar显色培养基和刀豆氨酸-甘氨酸-溴麝香草酚蓝(CGB)培养基35℃培养,使用API 20C AUX、全自动微生物鉴定仪Vitek 2 Compact、生物梅里埃基质辅助激光解析电离飞行时间质谱分析(MALDI-TOF MS)、r DNA ITS和IGS序列分析等方法对该菌进行菌种鉴定,并使用ATB FUNGUS 3进行抗真菌药敏试验。结果该菌在显色培养基上35℃培养48 h后为白色略带粉色菌落,在CGB培养基35℃培养5 d后使CGB培养基由黄色变为蓝色。API 20C AUX、Vitek 2 Compact和MALDI-TOF MS的鉴定结果均为新生隐球菌,r DNA ITS和IGS序列分析均提示该菌为格特隐球菌。5-氟胞嘧啶、两性霉素B、氟康唑、伊曲康唑、伏立康唑对该菌的最小抑菌浓度分别是≤4μg/m L、≤0.5μg/m L、≤1μg/m L、≤0.125μg/m L、≤0.06μg/m L。结论 CGB培养基、r DNA ITS和IGS序列分析能够区分新生隐球菌和格特隐球菌,应重视格特隐球菌的分离鉴定。
Objective To report a case of meningitis caused by Cryptococcus neoformans in Hebei Province, China. Methods Cryptococci isolated from cerebrospinal fluid of patients were respectively inoculated into Chromagar medium and Cn-glycine-glycine-glyceraldehyde (CGB) medium at 35 ℃ for culture. Using API 20C AUX, automatic microorganism analyzer Vitek 2 Compact, BioMérieux-MALDI-TOF MS, rDNA ITS and IGS sequence analysis were used to identify the bacteria and ATB FUNGUS 3 was used for antifungal susceptibility test. Results The strain was slightly pinkish white colonies after cultured at 35 ℃ for 48 h on the chromogenic medium. After cultured for 5 days at 35 ℃ in CGB medium, the medium was changed from yellow to blue. Cryptococcus neoformans was identified by API 20C AUX, Vitek 2 Compact and MALDI-TOF MS, respectively. Sequence analysis of rDNA ITS and IGS indicated that the strain was C. cryptococcosis. The minimum inhibitory concentrations of 5-fluorocytosine, amphotericin B, fluconazole, itraconazole and voriconazole were ≤4μg / mL, ≤0.5μg / mL, ≤1μg / mL, ≤ 0.125 μg / m L, ≤ 0.06 μg / m L. Conclusion CGB medium, rDNA ITS and IGS sequence analysis can distinguish Cryptococcus neoformans and Cryptococcus neoformans, should be focused on the identification of Cryptococcus.