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Measles virus RNA is not detected in inflammatory bowel disease using hybrid capture and reverse transcription followed by the polymerase chain reaction
Author(s) -
Chadwick Nicholas,
Bruce Ian J.,
Schepelmann Silke,
Pounder Roy E.,
Wakefield Andrew J.
Publication year - 1998
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/(sici)1096-9071(199808)55:4<305::aid-jmv9>3.0.co;2-4
Subject(s) - measles virus , virology , rna , biology , virus , morbillivirus , measles , reverse transcriptase , reverse transcription polymerase chain reaction , microbiology and biotechnology , paramyxoviridae , gene , gene expression , viral disease , vaccination , genetics
Recent epidemiological and immunohistochemical studies have indicated a possible link between measles virus and inflammatory bowel disease (IBD). The aim of this study was to use a sensitive and robust method for the detection of measles virus RNA in IBD and control clinical samples. Peripheral blood mononuclear cells and intestinal resection tissue from IBD and control patients were studied. Two methods were used to determine the presence of measles virus RNA: hybrid capture, using measles virus‐specific oligonucleotides linked to paramagnetic solid‐phase supports, was carried out on total cellular RNA to enrich for measles virus RNA sequences. Reverse transcription followed by the polymerase chain reaction (RT‐PCR) using rTth DNA polymerase was employed for amplification of measles virus N‐gene sequences amongst the enriched species. Total RNA was also used for RT‐PCR of a housekeeping mRNA species to assess RNA quality. RT‐PCR for another region of the measles genome (the haemagglutinin (H) gene) was also undertaken in order to confirm the results obtained using N‐gene primers for analysis of these samples. None of the samples were positive for measles N‐ or H‐gene RNA using RT‐PCR. Positive control samples confirmed the sensitivity of the methods employed. These results show that either measles virus RNA was not present in the samples, or was present below the sensitivity limits known to have been achieved. J. Med. Virol. 55:305–311, 1998 . © 1998 Wiley‐Liss, Inc.