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Journal of Clinical Microbiology, November 2005, p. 5509-5514, Vol. 43, No. 11
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.11.5509-5514.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Rapid Identification of Human Adenovirus Types 3 and 7 from Respiratory Specimens via Multiplex Type-Specific PCR

Jin A Lee,1 Nam Hee Kim,1 Sun Jung Kim,2 Eun Hwa Choi,1,2 and Hoan Jong Lee1*

Department of Pediatrics, Seoul National University College of Medicine,1 Medical Science Research Institute, Seoul National University Bundang Hospital, Seoul, Korea2

Received 31 January 2005/ Returned for modification 14 May 2005/ Accepted 27 August 2005

The rapid diagnosis of human adenovirus (Ad) infection is crucial for the timely recognition of epidemics. Moreover, identification of the serotypes known to cause serious disease can be helpful in therapeutic intervention. A multiplex PCR assay was developed for the rapid detection of adenovirus type 3 (Ad3) and Ad7 directly from clinical specimens. For this assay, three primer pairs (primers were based on the conserved and hypervariable regions of the hexon) were designed in order to simultaneously amplify all adenoviral serotypes and discriminate between Ad3 and Ad7. In our preliminary analysis, this multiplex PCR assay generated amplicons of the consensus primers from all 106 adenoviral isolates of diverse serotypes and proved able to correctly identify Ad3 and Ad7. This assay was subsequently applied to the detection of Ad3 and Ad7 in respiratory specimens. Among the 127 nasal aspirates from which an adenovirus was grown, the sensitivity with which any serotype could be detected was 91% (115/127). Two of the 53 nasal aspirates which did not grow Ads yielded adenovirus-specific bands, which were confirmed by sequencing analysis. Among the 115 specimens which produced common adenoviral bands, the sensitivity with which Ad3 could be detected was 93% (26/28), and the sensitivity with which Ad7 could be detected was 100% (35/35). Five out of the 115 specimens were proved to harbor more than one type of Ad via sequencing analysis of the amplicons, suggesting mixed infection with at least two different serotypes. In conclusion, this multiplex PCR system can be utilized in the rapid identification of Ad3 and Ad7 directly from clinical specimens. Furthermore, this method constitutes a diagnostic strategy for the detection of coinfection by different Ad serotypes.

* Corresponding author. Mailing address: Department of Pediatrics, Seoul National University Children's Hospital, 28 Yongon-dong, Chongno-gu, Seoul 110-744, Korea. Phone: 82-2-2072-3633. Fax: 82-2-745-4703. E-mail: hoanlee{at}snu.ac.kr.

Journal of Clinical Microbiology, November 2005, p. 5509-5514, Vol. 43, No. 11
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.11.5509-5514.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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