Comparative detection of enterovirus RNA in cerebrospinal fluid: GeneXpert system vs. real-time RT-PCR assay

Abstract

Enteroviruses (EVs) constitute the most common cause of aseptic meningitis in both children and adults. Molecular techniques have now been recognized as the reference standard for the diagnosis of EV infections, and the rapidity of the molecular diagnosis of EV meningitis has been shown to be a determining factor in the management of patients. The rapid documentation of EV RNA in cerebrospinal fluid (CSF) is key to adapting patient management and the therapeutic regimen. To shorten the time needed for virological documentation, we implemented EV RNA detection in two point-of-care (POC) laboratories. Here, we present the results of the POC detection of EV RNA with the Xpert EV kit on the GeneXpert integrated system, and a comparison with the real-time RT-PCR (rtRT-PCR) assay routinely used in the core virology laboratory. From January to September 2009, a total of 310 CSF samples were tested. The rtRT-PCR gave 81 positive, 225 negative and four 'indeterminate' results. POC results were concordant in 81.6% (253/310). Most of the discrepancies consisted of 'indeterminate' results at the POC level (16%). Calculated performances (excluding the indeterminate results) of the Xpert EV kit on the GeneXpert system in POC settings were 100%, 98.9%, 97.6% and 100% for Sensibility, Specificity, positive predictive value and negative predictive value, respectively. Taken together, these results indicate that the implementation of POC detection of EV RNA can provide robust results in <4 h, and may have a significant impact on patient management, therapeutic attitude, and hospitalization costs.

Impact of Enteroviral Polymerase Chain Reaction Testing on Length of Stay for Infants 60 Days Old or Younger

“…Commercially available EV PCR testing platforms with fully automated processes have the potential for test turnaround in as little as one hour. 25,26 However, in practice, test turnaround time typically ranges between 12 to 24 hours, reflecting limited testing hours and specimen batching. 7,10,13 As clinical EV PCR testing evolves, earlier availability of test results may increase the impact on LOS.…”

An Epidemic of Dengue-1 in a Remote Village in Rural Laos

“…Viral RNA was extracted using EZ1 Virus v2.0 Kit and EZ1 Biorobot (Qiagen) from 200 µl of acute serum following manufacturer's instruction and eluted with 90 µl of elution buffer. Internal phage controls were added to all sera before extraction to monitor the amplification processing and check for the presence of PCR inhibitors [16].…”

Investigation of the Causative Agents of Meningitis By Real-Time PCR Method

“…PCR çalış-ması ile 4 saatten daha kısa bir sürede hastalara sonuç verilir. Bu da klinisyenin daha erken tedaviye başlaması, hastanın erken iyileşmesi ve dolayısıyla yatış maliyetinin düşmesiyle sonuçlanır (29,30) .…”