Molecular Assays Detect Enterovirus in Clinical Samples

Different real-time polymerase chain reaction assays (PCR) assays and protocols for EV RNA detection are available, but comparisons of results obtained according to the different approaches are limited and many of these molecular assays are laborious and time-consuming.

Scientists at the Hospital Sant Joan de Déu (Barcelona, Spain) evaluated an in-house real-time reverse transcription (RT-PCR) assay with a commercialized TaqMan real-time RT-PCR kit for detection of EV. They tested and analyzed 132 patient samples between July and November 2010. These were consecutive clinical specimens from pediatric patients less than six years old with clinical suspicion of EV infection.

The optimized one-step RT-PCR amplification reactions were undertaken in a single reaction tube using the Enterovirus real-time RT-PCR microplate system (TaqMan) kit, MutaPLATE Enterovirus (Immundiagnostik AG; Bensheim, Germany). The in-house reaction of a one-step RT-PCR amplification was performed in a single reaction tube using the primers and probe synthesized by Applied Biosystems (Foster City, CA, USA).

A total of 153 samples from 132 patients were collected during the study period. There were 68 males (51.5%) and 64 females (48.5%); the age of patients ranged from 3 days to 72 months. Samples were collected from patients with suspicion of EV infection who presented fever without focus or viral meningitis. The samples tested included 101 (66%) plasmas, 51 (33.3%) cerebrospinal fluids (CSF), and 1 (0.7%) vesicle.

A total of 19 of 132 patients (14.4%) involving 20 samples (14 plasma and six CSF samples) were positive in at least one of the two assays. The sensitivity of the in-house assay when the MutaPLATE assay was used as a reference was 90% and the specificity was 100%. The authors concluded that the two real-time RT-PCR assays are rapid and easy methods for detection of EV and contribute positively to rational and specific management of the etiological cause of fever in children.

Enteroviruses are RNA viruses belonging to the Picornaviridae family. EV infections are common, especially in young children, and are associated with a multitude of clinical manifestations. The study was published in the May 2012 issue of the European Journal of Clinical Microbiology & Infectious Diseases.

Related Links:
Hospital Sant Joan de Déu
Immundiagnostik AG
Applied Biosystems