广西地区隐球菌感染的临床特征、菌种鉴定及体外抗真菌药物敏感性

In vitro antifungal susceptibility profiles of Cryptococcus neoformans and Cryptococcus gattii clinical isolates of in Guangxi,Southern China

目的分析广西地区隐球菌感染的临床特征、致病菌种鉴定及其体外药物敏感性。方法收集86例确诊隐球菌病患者(14例为HIV阳性,72例为HIV阴性)中分离出103株隐球菌及其患者资料并对其临床特征进行分析,使用MALDI-TOF MS和ITS区测序分子鉴定方法,92株被鉴定为新生隐球菌grubii变种,11株被鉴定为格特隐球菌。使用CLSI M27-A4方法对菌株进行常用抗真菌药物氟康唑、两性霉素B、5-氟胞嘧啶、伊曲康唑、伏立康唑、泊沙康唑和艾沙康唑体外药物敏感性测试。结果①86例患者(男性59名,女性27名),年龄为21~84岁,其中55人无基础疾病,31人有基础疾病。②由于目前对于隐球菌尚无泊沙康唑、艾沙康唑和两性霉素B的折点,因此参考了念珠菌属的数据,所有菌株对大多数抗真菌药物敏感。抗真菌药物对新生隐球菌grubii变种最低抑菌浓度范围为:氟康唑0.05~4μg/mL,两性霉素B 0.25~1μg/mL,5-氟胞嘧啶0.0625~2μg/mL,伊曲康唑0.0625~0.25μg/mL,伏立康唑0.0078~0.25μg/mL,泊沙康唑0.0313~0.5μg/mL,艾沙康唑0.0020~0.125μg/mL。抗真菌药物对格特隐球菌最低抑菌浓度范围为:氟康唑1~16μg/mL,5-氟胞嘧啶0.125~1μg/mL,两性霉素B 0.25~1μg/mL,伊曲康唑0.0625~0.25μg/mL,伏立康唑0.0156~0.125μg/mL,泊沙康唑0.0156~0.25μg/mL,艾沙康唑0.0078~0.125μg/mL。结论MALDI-TOF MS是一种快速可靠的鉴定隐球菌的方法。广西地区分离隐球菌对临床抗真菌药物敏感,抗真菌药敏试验有助于早期发现耐药菌株,对有效地治疗隐球菌病具有重要意义。

Objective To analyze the clinical characteristics,pathogenic strain identification and in vitro antifungal susceptibility of Cryptococcus neoformans and Cryptococcus gattii in Guangxi.Methods One hundred and three Cryptococcus strains were isolated from 86 patients with confirmed cryptococcosis(14 were HIV-positive and 72 were HIV-negative)and their clinical characteristics were analyzed.92 strains were identified as Cryptococcus neoformans grubii variant and 11 strains were identified as Cryptococcus gattii using MALDI-TOF MS and ITS region sequencing molecular identification methods.The strains were tested for in vitro antifungal susceptibility to the commonly used antifungal drugs including fluconazole,amphotericin B,5-fluorocytosine,itraconazole,voriconazole,posaconazole,and esaconazole referring to the CLSI M27-A4 method.Results There were 59 males and 27 females aged 21-84 years in 86 patients,55 of them had no underlying disease.Referring to the interpretation criteria in CLSI and EUCAST,all strains were sensitive to most antifungal drugs.The minimum inhibitory concentrations of antifungal drugs against Cryptococcus neoformans var.grubii ranged from 0.05-4μg/mL for fluconazole,0.25-1μg/mL for amphotericin B;0.0625-2μg/mL for 5-fluorocytosine,0.0625-0.25μg/mL for itraconazole,0.0078-0.25μg/mL for voriconazole,0.0313-0.5μg/mL for posaconazole,and 0.0020-0.125μg/mL for estaconazole.The minimum inhibitory concentrations of antifungal drugs against Cryptococcus gattii ranged from 1-16μg/mL for fluconazole,0.125-1μg/mL for 5-fluorocytosine,0.25-1μg/mL for amphotericin B,0.0625-0.25μg/mL for itraconazole,0.0156-0.125μg/mL for voriconazole,0.0156-0.25μg/mL for posaconazole,and 0.0078-0.125μg/mL for esaconazole.Conclusion MALDI-TOF MS is a rapid and reliable method for the identification of Cryptococcus.Cryptococci isolated in Guangxi are sensitive to clinical antifungal drugs,and the antifungal sensitivity test helps to detect drug-resistant strains at an early stage,which is important for the effective treatment of cryptococcosis.