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Journal of Clinical Microbiology, February 2005, p. 879-885, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.879-885.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Novel Replicon-Based Reporter Gene Assay for Detection of Rubella Virus in Clinical Specimens

Wen-Pin Tzeng,1 Yumei Zhou,1 Joseph Icenogle,2 and Teryl K. Frey1*

Department of Biology, Georgia State University,1 Measles Virus Section, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia2

Received 5 November 2003/ Returned for modification 16 February 2004/ Accepted 18 August 2004

Proof of concept for a novel diagnostic assay for rubella virus (RUB) based on RUB replicons expressing reporter genes was demonstrated. RUB replicons have the structural protein coding region replaced with a reporter gene such as green fluorescent protein or chloramphenicol acetyltransferase. Previously, it was shown that a replicon construct with a specific in-frame deletion in the nonstructural protein coding region (NotI, approximately nucleotides 1500 to 2100 of the genome) failed to replicate and express the reporter gene unless rescued by a coinfecting wild-type helper RUB (W.-P. Tzeng et al., Virology 289:63-73, 2001). In the present study, it was found that rescue of reporter gene expression by NotI replicons occurred when coinfection was done with clinical specimens containing RUB, indicating that this system could be the basis for a diagnostic assay. The assay was sensitive, using laboratory RUB strains and as low a dose as one plaque-forming unit. The assay was specific in that it was positive for RUB strains of both genotypes and was negative for a panel of human viruses. It was also possible to genetically sequence the RUB present in positive clinical specimens detected in the assay for genotypic strain determination.

* Corresponding author. Mailing address: Department of Biology, Georgia State University, 24 Peachtree Center Ave., Atlanta, GA 30303. Phone: (404) 651-3105. Fax: (404) 651-3105. E-mail: tfrey{at}gsu.edu.

Journal of Clinical Microbiology, February 2005, p. 879-885, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.879-885.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Zhu, Z., Xu, W., Abernathy, E. S., Chen, M.-H., Zheng, Q., Wang, T., Zhang, Z., Li, C., Wang, C., He, W., Zhou, S., Icenogle, J. (2007). Comparison of Four Methods Using Throat Swabs To Confirm Rubella Virus Infection. J. Clin. Microbiol. 45: 2847-2852 [Abstract] [Full Text]  
  • Mori, N., Motegi, Y., Shimamura, Y., Ezaki, T., Natsumeda, T., Yonekawa, T., Ota, Y., Notomi, T., Nakayama, T. (2006). Development of a new method for diagnosis of rubella virus infection by reverse transcription-loop-mediated isothermal amplification.. J. Clin. Microbiol. 44: 3268-3273 [Abstract] [Full Text]  


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