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Enrichment of cerebrospinal fluid samples on cell culture for enhancement of sensitivity of mumps and enterovirus detection by multiplex RT-PCR.
Diagn Microbiol Infect Dis. 2008 Apr; 60(4):375-9.DM

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.

Authors+Show Affiliations

Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, P.O. Box 14155-331, Tehran, Iran.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

18083320

Citation

Kermanian, Masoumeh, et al. "Enrichment of Cerebrospinal Fluid Samples On Cell Culture for Enhancement of Sensitivity of Mumps and Enterovirus Detection By Multiplex RT-PCR." Diagnostic Microbiology and Infectious Disease, vol. 60, no. 4, 2008, pp. 375-9.
Kermanian M, Soleimanjahi H, Arefian E, et al. Enrichment of cerebrospinal fluid samples on cell culture for enhancement of sensitivity of mumps and enterovirus detection by multiplex RT-PCR. Diagn Microbiol Infect Dis. 2008;60(4):375-9.
Kermanian, M., Soleimanjahi, H., Arefian, E., & Bamdad, T. (2008). Enrichment of cerebrospinal fluid samples on cell culture for enhancement of sensitivity of mumps and enterovirus detection by multiplex RT-PCR. Diagnostic Microbiology and Infectious Disease, 60(4), 375-9.
Kermanian M, et al. Enrichment of Cerebrospinal Fluid Samples On Cell Culture for Enhancement of Sensitivity of Mumps and Enterovirus Detection By Multiplex RT-PCR. Diagn Microbiol Infect Dis. 2008;60(4):375-9. PubMed PMID: 18083320.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enrichment of cerebrospinal fluid samples on cell culture for enhancement of sensitivity of mumps and enterovirus detection by multiplex RT-PCR. AU - Kermanian,Masoumeh, AU - Soleimanjahi,Hoorieh, AU - Arefian,Ehsan, AU - Bamdad,Taravat, Y1 - 2008/02/20/ PY - 2007/05/30/received PY - 2007/10/30/revised PY - 2007/10/31/accepted PY - 2007/12/18/pubmed PY - 2008/5/28/medline PY - 2007/12/18/entrez SP - 375 EP - 9 JF - Diagnostic microbiology and infectious disease JO - Diagn Microbiol Infect Dis VL - 60 IS - 4 N2 - 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. SN - 0732-8893 UR - https://www.unboundmedicine.com/medline/citation/18083320/Enrichment_of_cerebrospinal_fluid_samples_on_cell_culture_for_enhancement_of_sensitivity_of_mumps_and_enterovirus_detection_by_multiplex_RT_PCR_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0732-8893(07)00507-X DB - PRIME DP - Unbound Medicine ER -