12例播散型组织胞浆菌病病原学调查

A primary investigation on disseminated histoplasmosis in Hubei

目的 调查湖北地区播散型组织胞浆菌病 (PDH)的流行情况。方法 将分布在湖北省 8地市 12例原诊断为黑热病患者的骨髓片重新镜检 ,部分患者外周血及骨髓制片镜检 ;骨髓、外周血及肝脾组织接种梅里埃双相培养基 (MLI) ,获单个菌落后转种BHIB、SDA、CMA培养基置 2 5℃和35℃孵育。将骨髓、外周血或分离培养的酵母相组织胞浆菌 (H·cap)制成浓菌液腹腔接种昆明种小鼠和裸鼠 ,待其发病后取其脾组织镜检及培养 ,观察其特征。用菌丝相和酵母相H·cap接种尿素酶(URE)、明胶 (GEL) 2 5℃和 35℃培养 ;采用组织胞浆菌素皮下注射患者 ,观察反应 ;应用两性霉素B(AmB)治疗。结果 12例患者骨髓片中单核 巨噬细胞内未见桑葚状细胞团、腊肠状细胞 ,1例发现杜利体 ,11例真菌孢子的胞浆中无动基体 ,未发现横隔 ;H·cap的URE反应阳性 ,不液化GEL ,2 5℃孵育时为菌丝相 ,未见帚状枝及孢子链 ,发现特征性齿轮状大分生孢子 ,不产生红色色素 ;35℃孵育时为酵母相 ,未见腊肠状孢子及横隔 ,镜下可见酵母相孢子。昆明种小鼠和裸鼠的脾组织可分离培养出H·cap ,其与马尔尼菲青霉菌、杜波伊斯胞浆菌等双相型真菌的菌丝相和酵母相菌落特征、镜下形态及生化反应有不同特点。结论 湖北地区有PDH的散发流行 ,且东南部发现率高于西?

Objective To investigate the epidemiologic features of disseminated histoplasmosis (PDH) in Hubei province. Methods Bone marrow smears of 12 patients diagnosed as Kala-azer in Hubei province including 4 patients in Jingsan, 2 patients in Shashi and each 1 in Yichang, Jinmen, Zhongxiang, Luotian, Xianning and Guanghua respectively were re-examed under microscope. Peripheral blood and bone marrow smears of several patients were detected. After inoculated the bone marrow, peripheral blood, liver and spleen tissue of patients in MLI, the single colony was trans-inoculated in BHIB, SDA and CMA and incubated at 25℃ and 35℃. Bone marrow, peripheral blood and bacterial fluid of yeast-phase Histoplasma capsulatum ( H·cap ) were injected into the abdominal cavity of Kunming mice and nude mice. When symptoms and signs developed, the spleen tissue was separated, then observed under microscope and cultured. Mycelium-phase and Yeast-phase H·cap were inoculated in urase and gelatin medium, then incubated at 25℃ and 35℃. Histoplasmin was injected subcutaneously into patients, and then followed for 48-72 hours. Amphotericin B was selected to treat the PDH patients. Results Moriform cell cluster and sausage-shaped cell were not observed in mononuclear-macrophages in the bone marrow smears from 12 patients. Leishman-Donovan body was found only in one patient. There wasn't kinetoplast in the cellular plasm of spores in 11 patients and no transeptae was found. The reaction of H·cap to urease was positive and H·cap did not liquefy the gelatin. It appeared to be mycelium-phase at 25℃ but no penicillus and catenulate conidia was found. The characteristic denticle macroconidia was observed but produced red coloring matter. It also appeared to be yeast-phase at 35℃. Yeast-phase spores were observed under microscope. No sausage-shaped spore and transeptae were identified. H·cap could be acquired in the spleen tissue in Kunming mice and nude mice. Bacterium forms, characteristics under microscope and biochemical reaction of mycelium-phase and yeast-phase H·cap were different from some other kinds of dimorphic fungi such as Penicillium marneffei and Histoplasm duboisii etc. Conclusion There were scattered epidemics of PDH in Hubei province. The detection rate of PDH was higher in the southeast area then in the northwest area. The golden standards of clinic diagnosis were mycological culture and inoculation to animals. Amphotericin B was necommended as the first choice for treatment.