title: Comparison of real-time florescence quantitative PCR measurements of VAD1 mRNA with three conventional methods in diagnosis and follow-up treatment of Cryptococcus neoformans infection.
abstract: This study was to develop a real-time florescence quantitative PCR (RT-FQ-PCR) assay to measure virulence-associated DEAD-box RNA helicase (VAD1) mRNA from Cryptococcus neoformans and evaluate its potential use in diagnosis and follow-up treatment of C. neoformans meningitis (CNM). Cryptococcus neoformans was detected using RT-FQ-PCR, ink staining, fungal culturing and C. neoformans antigen detection in CNM compared with a normal control. VAD1 mRNA was measured in both acute and stable CNM patients. The sensitivity of RT-FQ-PCR (96%) is higher than ink staining (72%) and culture culturing (64%) (P<0.05, P<0.05 respectively), but its sensitivity is the same as antigen detection (96%, P>0.05). The levels of VAD1 mRNA in the acute and stable phase of a C. neoformans infection are 3.042±0.906 and 2.187±0.665 respectively (P<0.01). The levels of VAD1 mRNA are correlated to the numbers of C. neoformans, intracranial pressure and glucose concentration in cerebrospinal fluid (CSF; P<0.01, P<0.01 and P<0.05 respectively). The levels of expression of VAD1 mRNA in the group of patients who received an AmB/5-FC/FZC drug regimen decreased more than in patients taking a 5-FC/AmB or 5-FC/FCZ drug combination. Quantitative measurements of VAD1 mRNA are valuable and reliable in diagnosing C. neoformans infection and evaluating a therapy response.
authors: Department of Laboratory Medicine, the First Affiliated Hospital, Fuzhou, 350005 Fujian, China. ouqishui@163.com
authors: Ou
authors: O
authors: Qishui
authors: Jiang
authors: J
authors: Ling
authors: Lin
authors: L
authors: Ni
authors: Yang
authors: Y
authors: Bin
authors: Li
authors: L
authors: Wen
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000935:Antifungal Agents / Q000627:therapeutic use; D002648:Child; D003455:Cryptococcus neoformans / Q000235:genetics*; D053487:DEAD-box RNA Helicases / Q000235:genetics*; D005260:Female; D005656:Fungal Proteins / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D016919:Meningitis, Cryptococcal / Q000175:diagnosis* / Q000188:drug therapy; D008875:Middle Aged; D012333:RNA, Messenger / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D012015:Reference Standards; D015203:Reproducibility of Results; D055815:Young Adult
chemical_list: D000935:Antifungal Agents; D005656:Fungal Proteins; D012333:RNA, Messenger; D053487:DEAD-box RNA Helicases