Diagnosis of enteroviral meningitis by using PCR with a colorimetric microwell detection assay

Author:

Rotbart H A1,Sawyer M H1,Fast S1,Lewinski C1,Murphy N1,Keyser E F1,Spadoro J1,Kao S Y1,Loeffelholz M1

Affiliation:

1. Department of Pediatrics, University of Colorado School of Medicine, Denver.

Abstract

A 5-h, user-friendly PCR assay for the diagnosis of enteroviral meningitis was developed. Reverse transcription and amplification were performed in a one-step reaction using rTth polymerase. Carryover contamination was prevented with dUTP and uracil N-glycosylate. Detection was performed colorimetrically on a microwell titer plate. Sensitivity, specificity, positive predictive value, and negative predictive value were 94.7, 97.4, 94.7, and 97.4%, respectively.

Publisher

American Society for Microbiology

Subject

Microbiology (medical)

Reference13 articles.

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2. The clinical relevance of CSF viral culture. A two-year experience with aseptic meningitis in Rochester;Chonmaitree T.;New York. JAMA,1982

3. Hsiung G. D. 1973. Enteroviruses p. 54-67. In G. D. Hsiung (ed.) Diagnostic virology. Yale University Press New Haven Conn.

4. Detection of Chlamydia trachomatis in endocervical specimens by polymerase chain reaction;Loeffelholz M. J.;J. Clin. Microbiol.,1992

5. Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions;Longo M. C.;Gene,1990

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