Virus minikits 2

We retrospectively compared CSF testing and acyclovir use in a pre-MEP era (1/1/2007–1/22/2017) to a post-implementation era of MEP (1/23/2017–12/31/17) amongst children >60 days with a CSF specimen sent to the Children’s Hospital Colorado (CHCO) microbiology laboratory for bacterial culture or viral PCR testing. A subanalysis compared children with CSF testing HSV negative by single-plex PCR through both eras with those testing HSV negative on MEP. CSF HSV singleplex-PCR testing was available throughout both eras, conducted following extraction using Virus Minikits 2.0 (Qiagen, Valencia, CA) on the BioRobot EZ1, using the MultiCode-RTx HSV 1&2 Kit (Luminex, Austin, TX). Following implementation, MEP was performed on-demand 24 hours/day, 7 days/week. Electronic medical record clinical decision support recommended ordering MEP if providers had concern for more than one pathogen, and discouraged ordering MEP in immunocompetent children >2 months without encephalitis with <5 white blood cells (WBCs)/μL in CSF.
Demographic and clinical characteristics were compared using Wilcoxon Rank-Sum tests, Fisher’s exact tests and chi-squared tests for continuous and categorical variables. Significance was set at 0.05. R version 3.4.1 software (R Foundation for Statistical Computing, Vienna, Austria, http://www.R-project.org/) was used for analysis.

Throughout both study periods, the CHCO microbiology laboratory offered singleplex PCR testing for HSV-1 and 2 using the MultiCode-RTx HSV 1&2 Kit (Luminex, Austin, TX), EV using the Xpert EV kit on GeneXpert (EV; Cepheid, Sunnyvale, CA), and CLIA approved lab developed RT-PCR tests for HHV-6(24 (link)), CMV, VZV, and HPeV(25 (link)) following nucleic acid extraction and processing with Virus Minikits 2.0 (Qiagen, Valencia, CA) on the BioRobot EZ1. CSF was cultured on blood and chocolate agar plates, as well as in blood culture broth bottles (Plus Aerobic/F and PedsPlus/F [Becton Dickinson and Co, Sparks, MD]) incubated in a BacTec 9120/9240 automated system (Becton Dickinson and Co). MEP testing was performed in real-time on-demand 24 hours/day, 7 days/week with conditional testing occurring following CSF cell counts if WBC≥5uL. MEP was performed in a dedicated biosafety cabinet to reduce the risk of contamination. Positive results on MEP were repeated in duplicate before reporting to providers with discrepant results repeated a third time to adjudicate.