Simultaneous detection of multiple pathogens by multiplex PCR coupled with DNA biochip hybridization

Hsiang Yun Tung, Wei Chen Chen, Bor Rung Ou, Jan Ying Yeh, Yeong Hsiang Cheng, Ping Hua Tsng, Ming Hua Hsu, Ming Shiun Tsai, Yu Chuan Liang

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)


Traditional serological enzyme-linked immunosorbent assay (ELISA) is routinely used to monitor pathogens during quarantine in most animal facilities to prevent possible infection. However, the ELISA platform is a single-target assay, and screening all targeted pathogens is time-consuming and laborious. In this study, to increase sensitivity and to reduce diagnosis time for high-throughput processes, multiplex PCR and DNA biochip techniques were combined to develop a multi-pathogen diagnostic method for use instead of routine ELISA. Eight primer sets were designed for multiplex PCR to detect genes from seven targeted bacterial and viral pathogens. DNA–DNA hybridization was conducted on a biochip following the multiple PCR analysis. Using this method, a total of 24 clinical samples were tested, and the result showed that not only single infection but also co-infection by multi-pathogens can be detected. In conclusion, multiplex PCR coupled with a DNA biochip is an efficient method for detecting multi-pathogens in a reaction. This platform is a useful tool for quarantine services and disease prevention in animal facilities.

Original languageEnglish
Pages (from-to)186-195
Number of pages10
JournalLaboratory Animals
Issue number2
Publication statusPublished - 2018 Apr 1

All Science Journal Classification (ASJC) codes

  • Animal Science and Zoology
  • veterinary(all)

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