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Clinical Chemistry 53: 188-194, 2007. First published December 7, 2006; 10.1373/clinchem.2006.071720
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(Clinical Chemistry. 2007;53:188-194.)
© 2007 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

Microarray-in-a-Tube for Detection of Multiple Viruses

Quanjun Liu, Yunfei Bai, Qinyu Ge, Shixin Zhou, Tian Wen and Zuhong Lua

State Key Laboratory of Bioelectronics, Southeast University, Nanjing, People’s Republic of China.

aAddress correspondence to this author at: State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, People’s Republic of China. Fax 086-25-83793779; e-mail zhlu{at}seu.edu.cn.

Background: The detection of multiple viruses is important for pathogenic diagnosis and disease control. Microarray detection is a good method, but requires complex procedures for multiple virus detection.

Methods: We developed a novel PCR assay, the microarray-in-a-tube system, which integrates multiple PCR processes and DNA microarrays for multiple virus detection. A 5 x 5 oligonucleotide microarray for detecting 4 respiratory tract viruses (severe acute respiratory syndrome–associated coronavirus, influenza A virus, influenza B virus, and enterovirus) with inner controls was arranged on the inner surface of a specially designed Eppendorf cap with a flat, optically transparent window.

Results: We were able to perform all detection processes in the encapsulated system without opening the cap. The 4 viruses were successfully amplified by one-step reverse transcription–PCR in the encapsulated tube. After the PCR process, the microarray-in-a-tube was inverted, and the fluorescence-labeled PCR products were directly hybridized on the microarray. Hybridization signals were obtained with an ordinary fluorescent microscope. The sensitivity of the system for virus detection reached 102 copies/µL. With the help of inner controls, the system provided reliable results without false negatives and false positives.

Conclusions: The microarray-in-a-tube system is a rapid, labor-saving tool for multiple virus detection with several advantages, such as convenience, prevention of cross-contamination of the PCR products, and potential for multiple-gene detection.






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Copyright © 2007 by the American Association for Clinical Chemistry.