荚膜相关蛋白10与DEAD-box的RNA解旋酶基因联合检测诊断新生隐球菌性脑膜炎  

Combined detection of capsule associated protein 10 and virulenceassociated DEADbox RNA helicase 1 inthe clinical diagnosis of Cryptococcus neoformans meningitis

作　　者：林旎[1] 江凌[2] 杨滨[2] 李雯[2] 欧启水[2] 

机构地区：[1]福建医科大学医学检验系,福州350004 [2]福建医科大学附属第一医院检验科,福州350004

出　　处：《中华传染病杂志》2012年第9期529-531,共3页Chinese Journal of Infectious Diseases

基　　金：福建医科大学苗圃基金资助项目（2010MP047）;福建省卫生厅青年基金资助项目（20102-59）

摘　　要：目的建立定量检测新生隐球菌荚膜相关蛋白10（cAPl0）和DEAD-box的RNA解旋酶（VADl）基因的方法，比较两者单独及联合应用诊断新生隐球菌感染的价值。方法选择行脑脊液真菌培养、墨汁染色或隐球菌抗原检测，其中一项阳性的新生隐球菌性脑膜炎患者23例，以同期颅脑外伤患者为对照。以新生隐球菌标准株构建质粒标准品，建立实时荧光定量PCR（RT-FQ-PCR）体系，检测脑脊液中的CAPl0和VADl。并与墨汁染色、真菌培养、隐球菌抗原检测比较。卡方检验评价两者单独或联合应用的诊断价值。结果在23例隐球菌性脑膜炎患者中，RTFQ-PCR法检测CAPl0mRNA阳性22例，阳性率为95。6％，墨汁染色法阳性16例，阳性率为69．6％（X^=4．167，P〈0．05），真菌培养阳性15例，阳性率为65．2％（X^=5．143，P〈0．05），抗原检测法阳性21例，阳性率为91。3％（X^=20．500，P〉0。05）。联合检测CAPl0及VADl基因诊断新生隐球菌性脑膜炎与单独检测cAPl0或VADl比较，差异无统计学意义（P〉0．05）。结论成功建立以新生隐球菌毒力基因为靶标的RT-FQ-PCR检测方法，检测结果优于传统方法。establish the quantitative detection of capsule associated protein 10 （CAP10） and virulence-associated DEAI〉box RNA helicase 1 （VAD1） genes in Cryptococcus neoforrnans （CN） and compare the diagnostic values of single gene test and combined gene test in CN meningitis. Methods Twenty-three CN meningitis patients with fungal culture or ink staining or CN antigen detection positive in cerebrospinal fluid （CSF） were recruited and patients with craniocerebral trauma were recruited as controls. Standard plasmids were constructed using standard CN strain. Real time fluorescent quantitative polymerase chain reaction （RT-FQ-PCR） was established to detect the mRNA expressions of CAP10 and VAD1 genes in the CSF of patients with CN meningitis, which were compared with the results of CSF ink staining, fungal culture and antigen detection. The diagnostic values of single gene test and combined gene test were compared by chi square test. Results Among 23 CN meningitis patients, 22 （95.6%） were CAP10 mRNA positive detected by RT-FQ-PCR, which was significantly higher than both ink staining （16/23, 69.6%, Z2 = 4. 167, P〉O. 05） and fungal culture （15/23, 65. 2%0, X2 =5. 143, P%0. 05）, respectively; but not significant different fromantigen detection （21/23, 91.3%, X^2 =0. 500, P2〉0.0S）. There were also no statistical significant differences between combined detection of CAP 10＋VAD1 and CAP 10 or VAD1 single gene test （P〉0.05）. Conclusion RT-FQPCR detection is successfully established using virulence genes as target, which is superior to the conventional methods.

关 键 词：隐球菌 新型 细菌荚膜 真菌蛋白质类 实时聚合酶链反应 RNA 信使 

分 类 号：R519[医药卫生—内科学]