Enrichment of cerebrospinal fluid samples on cell culture for enhancement of sensitivity of mumps and enterovirus detection by multiplex RT-PCR

Abstract

It has been demonstrated that the detection of enteroviruses and mumps virus nucleic acid in cerebrospinal fluid (CSF) specimens improves the management of the patients with aseptic meningitis. To determine the effect of overnight enrichment of mumps and enteroviruses in CSF samples on cell culture for increasing the sensitivity of viral detection, we developed a multiplex reverse transcriptase-polymerase chain reaction (RT-PCR). CSFs were collected from 100 patients younger than 7 years. The samples were tested for the presence of enteroviruses and mumps virus by multiplex RT-PCR method. Negative samples in Multiplex RT-PCR were enriched for viral template RNAs by overnight culture of CSF samples on cells and followed by the optimized Multiplex RT-PCR. Overall, 35% of the CSF samples were positive for enteroviruses, whereas only 1% of CSF samples were positive for both viruses. After enriching in cell culture, 34% of the negative samples showed a positive polymerase chain reaction band for enteroviruses, and a 3% increase was observed for both viruses. The results showed that enrichment of viral template RNAs in cell culture can increase the sensitivity of the multiplex RT-PCR assay and provide a rapid and sensitive method for detection of viral infections.