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Journal of Clinical Microbiology, August 2008, p. 2652-2658, Vol. 46, No. 8
0095-1137/08/$08.00+0     doi:10.1128/JCM.00192-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Sensitive and Broadly Reactive Reverse Transcription-PCR Assays To Detect Novel Paramyxoviruses{triangledown}

Suxiang Tong,* Shur-Wern Wang Chern, Yan Li, Mark A. Pallansch, and Larry J. Anderson

Division of Viral Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333

Received 30 January 2008/ Returned for modification 13 April 2008/ Accepted 16 June 2008

We have developed a set of reverse transcription-PCR assays for the detection and identification of known and novel paramyxoviruses in clinical specimens. Primers were designed from the conserved motifs of the polymerase pol gene sequences to detect members of the Paramyxovirinae or Pneumovirinae subfamily or groups of genera within the Paramyxovirinae subfamily. The consensus-degenerate hybrid oligonucleotide primer design and seminested or nested PCR assay design were used to enhance the breadth of reactivity and sensitivity of the respective assays. Using expressed RNA and 10-fold dilution series of virus-infected tissue culture isolates from different members of the family or genera, these assays were able to detect on average between 100 and 500 copies of template RNA. The assays were specific to the respective group of genera or subfamily viruses. This set of primers enhances our ability to look for novel viruses in outbreaks and diseases of unknown etiology.

* Corresponding author. Mailing address: Gastroenteritis and Respiratory Viruses Laboratory Branch, 1600 Clifton Road, MS G18, Atlanta, GA 30333. Phone: (404) 639-1372. Fax: (404) 639-4005. E-mail: sot1{at}cdc.gov

{triangledown} Published ahead of print on 25 June 2008.

Journal of Clinical Microbiology, August 2008, p. 2652-2658, Vol. 46, No. 8
0095-1137/08/$08.00+0     doi:10.1128/JCM.00192-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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