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A sensitive and robust method for measles RNA detection

Chadwick, N., Bruce, Ian J., Davies, M., Van Gemen, B., Schukkink, R., Khan, K., Pounder, R., Wakefield, A. (1998) A sensitive and robust method for measles RNA detection. Journal of Virological Methods, 70 (1). pp. 59-70. ISSN 0166-0934. (doi:10.1016/S0166-0934(97)00168-7) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided. (Contact us about this Publication)
Official URL
http://dx.doi.org/10.1016/S0166-0934(97)00168-7

Abstract

The aim of this study was to compare measles RNA amplification methods and to develop and select the most rapid, sensitive and robust procedure. The use of hybrid capture for measles RNA isolation was evaluated, and three RNA amplification detection techniques wen compared. These were: (a) reverse transcription followed by nested polymerase chain reaction (RT-PCR) with MMLV reverse transcriptase and Taq polymerase; (b) a combined RT-PCR reaction using rTth polymerase; and (c) NASBA. An internal positive control was also developed. The sensitivities of the detection methods were quantified by using a dilution series of a known amount of total RNA from measles-infected Vero cells or by calculation of the number of transcript molecules (produced from a recombinant plasmid containing an insert measles nucleoprotein DNA) present in each amplification reaction, respectively. The results indicated that hybrid capture followed by combined RT-PCR with rTth polymerase was the most reproducibly robust and sensitive protocol and could detect as few as 104 synthetic measles RNA transcripts added to tissue homogenates. However, NASBA proved to be the most sensitive method for measles RNA detection in water.

Item Type: Article
DOI/Identification number: 10.1016/S0166-0934(97)00168-7
Uncontrolled keywords: virus rna, accuracy; animal cell; article; controlled study; in vitro study; measles; measles virus; nonhuman; priority journal; quantitative assay; recombinant plasmid; reverse transcription polymerase chain reaction; rna analysis; technique; vero cell; virus infection, Animals; Blotting, Southern; Cercopithecus aethiops; DNA Primers; Electrophoresis, Agar Gel; False Negative Reactions; Humans; Measles virus; Nucleic Acid Amplification Techniques; Polymerase Chain Reaction; Reproducibility of Results; RNA, Viral; RNA-Directed DNA Polymerase; Sensitivity and Specificity; Vero Cells
Divisions: Faculties > Sciences > School of Physical Sciences
Faculties > Sciences > School of Physical Sciences > Functional Materials Group
Depositing User: I. Bruce
Date Deposited: 31 Jan 2019 07:27 UTC
Last Modified: 30 May 2019 08:49 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/71980 (The current URI for this page, for reference purposes)
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