两种血清隐球菌荚膜多糖抗原检测方法在肺隐球菌病中的应用研究

中国真菌学杂志　 2019, Vol. 14 　Issue (5): 264-269.

两种血清隐球菌荚膜多糖抗原检测方法在肺隐球菌病中的应用研究

黄进宝¹, 李红艳¹, 兰长青², 吕骁¹, 林志来¹, 王新航¹, 张宏英¹, 邹盛华³, 翁恒¹

1. 福建医科大学教学医院福建省福州肺科医院呼吸内科, 福州 350008;
2. 福建医科大学教学医院福建省福州肺科医院影像科, 福州 350008;
3. 福建医科大学教学医院福建省福州肺科医院检验科, 福州 350008

收稿日期:2019-01-31 出版日期:2019-10-28 发布日期:2019-10-28
通讯作者: 翁恒,E-mail:wengeng@sina.com E-mail:wengeng@sina.com
作者简介:黄进宝,男,硕士,主治医师.E-mail:huangnbao0@126.com
基金资助:
福建省临床重点专科建设项目（闽卫医政函[2018]145号）；福建省福州市临床医学中心项目资助（2018080305）

Clinical application and evaluation oftwo different methods for detection of cryptococcal capsular polysaccharide antigen in diagnosis and treatment of pulmonary cryptococcosis

HUANG Jin-bao¹, LI Hong-yan¹, LAN Chang-qing², LV Xiao¹, LIN Zhi-lai¹, WANG Xin-hang¹, ZHANG Hong-ying¹, ZOU Sheng-hua³, WEN Heng¹

1. Department of Respiratory Medicine, Fuzhou Pulmonary Hospital of Fu Jian, Educational Hospital of Fujian Medical University, Fuzhou 350008, China;
2. Department of Radiology and Imaging Sciences, Fuzhou Pulmonary Hospital of Fu Jian, Educational Hospital of Fujian Medical University, Fuzhou 350008, China;
3. Department of Laboratory Medicine, Fuzhou Pulmonary Hospital of Fu Jian, Educational Hospital of Fujian Medical University, Fuzhou 350008, China

Received:2019-01-31 Online:2019-10-28 Published:2019-10-28

摘要/Abstract

摘要：

目的探讨并比较血清酶联免疫分析法（ELISA）和侧流免疫层析法（LFA）检测血清隐球菌荚膜多糖抗原（CrAg）在肺隐球菌病（PC）中的应用价值。方法采集2016年1月~2018年6月福建省福州肺科医院109例疑似PC患者的血液，采用ELISA和LFA两种方法对血清CrAg进行检测，对比分析两种方法对PC的诊断效率，同时监测治疗过程中血清CrAg的浓度变化。结果 109例患者中，经肺活检或肺穿刺液隐球菌检测确诊为PC53例，非PC56例，检验结果如下：（1）应用ELISA方法，PC组血清CrAg浓度为[11.43（5.92，47.96）]μg/L，其中CrAg ≤ 3.2 μg/L的例数明显低于非PC组，而CrAg ≥ 5.0 μg/L的例数则明显高于非PC组，两者比较差异均有统计学意义（均P<0.05）。应用LFA方法，PC组血清CrAg检测阳性率明显高于非PC组，两者比较差异有统计学意义（P<0.05）。（2）应用ROC曲线分析显示，ELISA检测血清CrAg的曲线下面积为0.939（95%可信区间为0.892~0.985）。其中截断值取3.54 μg/L时约登指数最高，视为最佳截断值，对应的敏感度、特异度、阳性预测值及阴性预测值分别为94.3%、80.4%、82.0%和93.8%。（3）当血清CrAg取4.0 μg/L为截断值时，ELISA和LFA的诊断敏感度（88.7%和83.0%）两者比较差异无统计学差异（P>0.05），但ELISA特异度（82.1%）明显低于LFA（98.2%），两者比较差异有统计学意义（P<0.05）。当截断值分别取5.0 μg/L、6.0 μg/L和7.0μg/L时，ELISA和LFA的诊断敏感度及特异度相当（均P>0.05）。当取8.0 μg/L为截断值时，ELISA的诊断敏感度（62.3%）明显低于LFA（83.0%），两者比较差异有统计学意义（P<0.05），但特异度（100%和98.2%）两者大致相仿（P>0.05）。（4）14例患者在抗真菌治疗过程中监测血清CrAg的浓度变化，治疗前荚膜抗原浓度为[19.33（7.11，43.46）]μg/L，治疗后6个月荚膜抗原浓度为[8.09（5.39，11.90）]μg/L，较前明显下降，两者比较差异有统计学意义（P<0.05）。结论血清CrAg浓度为3.54 μg/L时视为ELISA的最佳截断值，但当取5.0 μg/L为截断值时，ELISA同时有较好的诊断敏感度和特异度，且和LFA的诊断效率相当，两种方法均有助于对PC的快速诊断，值得临床推广。与LFA定性检测相比，ELISA可动态监测血清CrAg的浓度变化，有助于PC的疗效判断和预后评估。

关键词: 侧流免疫层析, 酶联免疫, 隐球菌荚膜多糖抗原, 肺隐球菌病

Abstract:

Objective To evaluate lateral flow assay (LFA) and enzyme-linked immunesorbent assay (ELISA) for detection of cryptococcal capsular polysaccharide antigen (CrAg) in diagnosis and treatment of pulmonary cryptococcosis (PC). Method One hundred and nine cases of suspected PC were collected from January 2016 to June 2018 at Fuzhou Pulmonary Hospital of Fujian.Serum CrAg was detected by ELISA and LFA at the same time, and the diagnostic efficiency of the two methods for PC was compared and analyzed. Serum CrAg concentration was also monitored by ELISA during the treatment. Results Among the 109 patients, 53 cases were confirmed as PC by lung biopsy or cryptococcus detection of lung puncture liquid,56 cases were other lung diseases (Non-PC). The results were as follows:(1) The serum CrAg concentration detected by ELISA in the PC group was[11.43 (5.92, 47.96)] μg/L. The number of cases with serum CrAg ≤ 3.2 μg/L in the PC group was significantly lower than that in the non-PC group (P<0.05), while the number of cases with serum CrAg>5.0 μg/L was significantly higher than that in the non-PC group (P<0.05). The positive rate of serum CrAg detected by LFA in the PC group was significantly higher than that in the non-PC group, and the difference was statistically significant (P<0.05). (2) Receiver Operating Characteristic (ROC) curve analysis showed that the area under the curve of serum CrAg detected by ELISA was 0.939 (95% confidence interval was 0.892~0.985), and 3.54 μg/L was considered as the best cut-off value with the highest Youden index. The sensitivity, specificity, positive predictive value and negative predictive value were 94.3%, 80.4%, 82.0% and 93.8%, respectively. When 5.0 μg/L was taken as the diagnostic cut-off value of serum CrAg, there was no statistical difference in the diagnostic sensitivity between ELISA and LFA (88.7% vs 83.0%, P>0.05), but the specificity of ELISA test (82.1%) was significantly lower than that of LFA test (98.2%), and the difference was statistically significant (P<0.05). The diagnostic sensitivity and specificity of ELISA and LFA were similar (P>0.05) when the cut-off values of serum CrAg were 5.0μg/L, 6.0μg/L and 7.0 μg/L, respectively. The diagnostic sensitivity (62.3%) of ELISA was significantly lower than that of LFA (83.0%) when 8.0 μg/L was taken as the cut-off value of serum CrAg (P<0.05), but the specificity (100% and 98.2%) in the two methods was the same (P>0.05). (4) The serum CrAg concentration was continually monitored in 14 patients during the antifungal therapy. After six months of treatment, the serum CrAg was significantly decreased[19.33 (7.11, 43.46) vs 8.09 (5.39, 11.90) μg/L, P<0.05]. Conclusions 3.54 μg/L was considered as the best diagnostic cut-off value of serum CrAg of ELISA test. However, when 5.0 μg/L was taken as the diagnostic cut-off value, ELISA had good diagnostic sensitivity and specificity similarly to those of LFA, and the two methods had the same diagnostic efficiency. ELISA and LFA were both helpful to the rapid diagnosis of PC and worthy of being widely applied in clinic. Compared with LFA qualitative detection, ELISA could dynamically monitor the changes of serum CrAg concentration, which was useful for the treatment response and prognosis evaluation of PC.

Key words: lateral flow assay, enzyme-linked immunosorbent assay, cryptococcal capsular polysaccharide antigen, pulmonary cryptococcosis

中图分类号:

R379.5

黄进宝, 李红艳, 兰长青, 吕骁, 林志来, 王新航, 张宏英, 邹盛华, 翁恒. 两种血清隐球菌荚膜多糖抗原检测方法在肺隐球菌病中的应用研究[J]. 中国真菌学杂志, 2019, 14(5): 264-269.

HUANG Jin-bao, LI Hong-yan, LAN Chang-qing, LV Xiao, LIN Zhi-lai, WANG Xin-hang, ZHANG Hong-ying, ZOU Sheng-hua, WEN Heng. Clinical application and evaluation oftwo different methods for detection of cryptococcal capsular polysaccharide antigen in diagnosis and treatment of pulmonary cryptococcosis[J]. Chinese Journal of Mycology, 2019, 14(5): 264-269.

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