Combined detection of capsule associated protein 10 and virulence-associated DEAD-box RNA helicase 1 in the clinical diagnosis of Cryptococcus neoformans meningitis
10.3760/cma.j.issn.1000-6680.2012.09.004
- VernacularTitle: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
- Author:
Ni LIN
;
Ling JIANG
;
Bing YANG
;
Wen LI
;
Qishui OU
- Publication Type:Journal Article
- Keywords:
Cryptococcus neoformans;
Bacterial capsules;
Fungal proteins;
Real-time polymerase chain reaction;
RNA,messenger
- From:
Chinese Journal of Infectious Diseases
2012;30(9):529-531
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish the quantitative detection of capsule associated protein 10 (CAP10)and virulence-associated DEAD-box RNA helicase 1(VAD1)genes in Cryptococcus neoformans (CN) and compare the diagnostic values of single gene test and combined gene test in CN meningitis.MethodsTwenty-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％,x2 =4.167,P＜0.05) and fungal culture (15/23,65.2％,x2=5.143,P＜0.05),respectively; but not significant different from antigen detection (21/23,91.3％,x2=0.500,P＞0.05).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).ConclusionRT-FQ-PCR detection is successfully established using virulence genes as target,which is superior to the conventional methods.
