Evaluation of a rapid and completely automated real-time reverse transcriptase PCR assay for diagnosis of enteroviral meningitis.

Nucleic acid amplification tests (NAATs) for enterovirus RNA in cerebrospinal fluid (CSF) have emerged as the new gold standard for diagnosis of enteroviral meningitis, and their use can improve the management and decrease the costs for caring for children with enteroviral meningitis. The Xpert EV assay (Cepheid, Sunnyvale, CA) is a rapid, fully automated real-time PCR test for the detection of enterovirus RNA that was approved by the U.S. Food and Drug Administration for in vitro diagnostic use in March 2007. In this multicenter trial we established the clinical performance characteristics of the Xpert EV assay in patients presenting with meningitis symptoms relative to clinical truth. Clinical truth for enteroviral meningitis was defined as clinical evidence of meningitis, the absence of another detectable pathogen in CSF, and detection of enterovirus in CSF either by two reference NAATs or by viral culture. A total of 199 prospectively and 235 retrospectively collected specimens were eligible for inclusion in this study. The overall prevalence of enteroviral meningitis was 26.04%. The Xpert EV assay had a sensitivity of 94.69% (90% confidence interval [CI] = 89.79 to 97.66%), specificity of 100% (90% CI = 99.07 to 100%), positive predictive value of 100%, negative predictive value of 98.17, and an accuracy of 98.62% relative to clinical truth. The Xpert EV assay demonstrated a high degree of accuracy for diagnosis of enteroviral meningitis. The simplicity and on-demand capability of the Xpert EV assay should prove to be a valuable adjunct to the evaluation of suspected meningitis cases.

National disease burden of respiratory viruses detected in children by polymerase chain reaction.

BACKGROUND: The epidemiology of community-acquired respiratory viruses at the Children's Hospital of Wisconsin between 1996 and 1998 was examined with molecular [multiplex (M) PCR] and standard virologic methods. METHODS AND RESULTS: A total of 3325 patients with lower respiratory infection (LRI) [bronchiolitis (42%), pneumonia (38%) and croup (12%)] were identified. It is estimated that 545,000 LRI hospitalizations occur each year in the United States in children younger than 18 years old (viral, 428,000; pneumonia, 221,000; bronchiolitis, 222,000; croup, 65,000), including a continued increase in bronchiolitis hospitalizations (47.8/1000; <1 year). The most common viruses detected each year in hospitalized children included respiratory syncytial virus (A and B; 117,000), parainfluenza virus (PIV-1 and -2, 48 000; PIV-3, 18,000) and influenza (A and B, 39,000). With multiplex reverse transcription polymerase chain reaction (MPCR), different patient populations demonstrated different LRI epidemiology in relation to specific syndromes and viral causes. At least 21% of LRI hospitalizations were in children with significant medical problems.PIVs were detected much more frequently in immunocompromised children than in previously healthy children (33% vs. 16%). Similarly 15% of respiratory syncytial virus was detected between May and October, mostly in immunocompromised children. Influenza caused 19% of bronchiolitis in previously healthy children, and no virus could be reliably predicted based on clinical presentation, including wheezing. Of children hospitalized for LRI, 40% tested positive for the seven most common respiratory viruses by MPCR. CONCLUSIONS: Work is under way to expand the number of viruses detected by MPCR and to determine whether newly discovered viruses, such as human metapneumovirus, contribute to the burden of pediatric LRI hospitalizations.

Mycoplasma hominis and Ureaplasma species brain abscess in a neonate.

We report an infant with a mixed Mycoplasma and Ureaplasma brain abscess who was successfully treated with intravenous doxycycline and erythromycin. Therapeutic concentrations of doxycycline were demonstrated in cerebrospinal fluid. This report is evidence of the potential for Mycoplasma and Ureaplasma to produce focal central nervous system infection, as well as meningitis, in neonates and that use of doxycycline can be efficacious in the therapy of such infections.