摘要
目的 探讨兔肺隐球菌感染模型制作方法及隐球菌荚膜多糖 （Crag）胶体金免疫层析法 （LFA）对肺隐球菌感染的诊断价值。方法 将36只新西兰大白兔随机分为免疫抑制组20只,免疫正常组10只和对照组6只。采用气管穿刺法对免疫抑制组及免疫正常组进行隐球菌菌种接种,对照组气管内注射生理盐水,免疫抑制组于实验第1-5天每日静脉注射阿糖胞苷,之后隔日1次维持免疫抑制状态;第4日起,每日1次万古霉素、头孢他定静脉注射,隔日1次庆大霉素静脉注射,预防细菌感染。免疫正常组及对照组未注射免疫抑制剂及抗生素。接种1周后抽取静脉血行CragLFA、1-3-β-D-葡聚糖检测 （G试验）、半乳甘露聚糖 （GM试验）,并行胸部CT扫描,之后处死取肺灌洗液行隐球菌荚膜抗原胶体金试验、肺组织进行病原菌培养、肺组织病理检查。结果 胸部CT检查免疫抑制组阳性率75% （15/20）,免疫正常组阳性率20% （2/10）,而且免疫抑制组出现磨玻璃影及斑片影、血管支气管束增粗、结节等多种表现,免疫正常组主要表现为结节影;隐球菌培养免疫抑制组阳性率100% （20/20）,免疫正常组阳性率30% （3/10）;病理检查免疫抑制组阳性率80% （16/20）,免疫正常组阳性率30% （3/10）;空白对照组6只兔子全部检查均阴性。免疫抑制组新型隐球菌组模型建立成功率为80 00% （16/20）,免疫正常组的成功率为30 00% （3/10）,两组差异有统计学意义 （P 〈0.05）;血清CragLFA敏感度为89 47%,特异度为100%;灌洗液CragLFA敏感度为100%,特异度为82 35%。G试验及GM试验均为阴性。结论 免疫功能抑制宿主更容易感染肺隐球菌。血清及肺泡灌洗液CragLFA对诊断肺隐球菌病均有较高的应用价值,特别是两者联合应用时。G试验、GM试验在肺隐球菌感染中多为阴性。
Objective To build of an animal model of pulmonary cryptococcosis infection and assess the value of the cryptococcal antigen （Crag） lateral flow assay （LFA）, 1-3-beta-d-glucan test （C-test）, glactomannan test （GM-test） to pulmonary cryptococcosis. Methods Thirty-six adult New Zealand whiterabbits were randomly divided into the three group： immunosuppressed group （n = 20 ）, immunocompetent group （n = 10 ） and control group （n = 6 ）. The immunosuppressed and immunoeompetent group inoculate soliquoid of cryptococcus neoformans via trachea by tracheal intubation. The control group injected the same dose of saline by the same way. In the immunosuppressive group, cytarabine was iniected intravenously on the day of 1-5, then maintained immunosuppressive state one time every other day ; On the fourth day, vancomycin and cephalosporin were given one time a day, and one time gentamicin was administered intravenously every other day to prevent bacterial infection. Immunocompetent group and control group animals didn＇t inject any drug. Then, CT scan was performed at one week after inoculation or before execution. We draw venous blood from the fringe of ear for cryptococcal antigen lateral flow assay, G-test and GM-test before the rabbits killed. Getting lung from rabbits for tissue culture, tissue pathology and bronehoalveolar lavage fluid（BALF）. Cryptococcal antigen concentration of BALF was detected by lateral flow assay. Results The positive rate of Chest CT was 75%（15/20） in immunosuppressed group and 20% （2/10） in immunocompetent group, meanwhile the immunosuppressed group showed a variety of manifestations, such as ground glass and porphyry shadow, thickening of the bronchobronchial bundles, and nodules, and the main presence of immunocompetent group was nodules. All immunosuppressed rabbits＇ lung tissue culture showed presence of eryptocoecus neoforman, while immunocompetent group showed two three of ten presence of cryptococcus neoforman. Pathological changes were detected in 80G （16/20） of immunosuppressed group, and in 30G（3/10） of immunocompetent group. Achievementratio of modeling in immunosuppressed group was 80%and in immunocompetent group was 30%, All tests in the control group were negative. The specificity of serum and BALF of the lateral flow assay were 100 %and 82.35 % respectively, meanwhile the sensitivity were 89.47%and 100%. It showed significant difference between the two groups. All of the results of G-test and GM-test were negative. Conclusions Immunosuppressed individuals are more susceptible to pulmonary eryptoeoecosis infection than those who are healthy individuals. The diagnostic value of lateral flow assay, both in serum and BALF, are very high both sensitivity and specificity in pulmonary cryptococcosis, especially when the two used together. G test and GM-test are negative in pulmonary cryptococeosis.
作者
李小华
赖国祥
许昌声
包宇旺
林新
陈文
方跃华
蔡立志
余天兴
胡辛兰
徐礼裕
Li Xiaohua;Lai Guoxiang;Xu Changsheng;Bao Yuwang;Lin xin;Chen Wen;Fang Yuehua;Cai Lizhi;Yu Tianxing;Hu Xinlan;Xu Liyu.(Department of Respiratory Medicine, the Fuzhou First Hospital Affiliated of Fujian Medical University, Fuzhou 350009, Chin)
出处
《国际呼吸杂志》
2018年第10期725-730,共6页
International Journal of Respiration
基金
福建省医学创新基金（2014-CX33）
福建省自然科学基金（2015J01497）
福州市临床重点专科项目[榕卫医（2016）57号]
关键词
隐球菌
肺隐球菌病
动物模型
隐球菌荚膜多糖胶体金免疫层析法
G试验
GM试验
Cryptococcus
Pulmonary cryptococcosis
Animal model
Cryptococcal antigen lateralflow assay
1-3 beta-d-glucan test
Glactomannan test
