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Two different PCR assays to detect enteroviral RNA in CSF samples from patients with acute aseptic meningitis
- Source :
- Journal of Medical Virology. 47:378-385
- Publication Year :
- 1995
- Publisher :
- Wiley, 1995.
-
Abstract
- Two polymerase chain reaction (RT-PCR) assays were developed to allow rapid detection of enteroviral RNA in cerebrospinal fluid samples (CSF). Primers homologous to the conserved 5′ noncoding region of the enterovirus genome were designed. The RT-PCR product size was-500 bp (479 bp for Poliovirus, 500 bp for Cox-sackievirus) and was visualized using ethidium bromide-stained gels. Assay 1 utilized Moloney Murine Leukaemia Virus Reverse Transcriptase (MMLV-RTase) for reverse transcription and Taq polymerase for subsequent PCR. Assay 2 utilized a thermoactive DNA polymerase of Thermus thermophilus (rTth enzyme) for both reverse transcription and DNA amplification. In addition, in Assay 2 reverse transcription and PCR wer eaccomplished within the same reaction tube. Both assays detected between 1 and 0.02 TCID50 of prototype strains of Polio and Coxsackie type B viruses propagated in VERO cell and spiked in a pooled preparation of CSF samples from patients with noninfective neurological disorders. However, Assay 1 was 10-fold more sensitive than Assay 2 when applied to the detection of enteroviral RNA in CSF samples from patients with etiologically well characterized acute aseptic meningitis. © Wiley-Liss, Inc.
- Subjects :
- Adult
DNA polymerase
Molecular Sequence Data
medicine.disease_cause
Polymerase Chain Reaction
Cell Line
law.invention
Feces
chemistry.chemical_compound
law
Virology
Chlorocebus aethiops
Tumor Cells, Cultured
medicine
Animals
Humans
Meningitis, Aseptic
Child
Vero Cells
Polymerase chain reaction
Enterovirus
Base Sequence
biology
Aseptic meningitis
RNA
RNA-Directed DNA Polymerase
medicine.disease
Reverse transcriptase
Infectious Diseases
Real-time polymerase chain reaction
chemistry
Child, Preschool
Acute Disease
DNA, Viral
biology.protein
RNA, Viral
Taq polymerase
Subjects
Details
- ISSN :
- 10969071 and 01466615
- Volume :
- 47
- Database :
- OpenAIRE
- Journal :
- Journal of Medical Virology
- Accession number :
- edsair.doi.dedup.....a93b25b67929386e38eb846092045fbd