Development of Two Multiplex Real-Time PCR Assays for the Rapid Detection of RNA and DNA Viruses Associated with Gastroenteritis in Pediatric Patients
Dongmei Chen1, Yu Sun1, Linqing Zhao1*, Yuan Qian1, Runan Zhu1, Liying Liu1, Liping Jia1, Huijin Dong1, Jie Deng1 and Li Deng2
1Laboratory of Virology, Capital Institute of Pediatrics, China
2Affiliated Children’s Hospital to Capital Institute of Pediatrics, China
- *Corresponding Author:
- Dr. Linqing Zhao
Associate Chief of Laboratory of Virology
Capital Institute of Pediatrics
Beijing, 100020, China
E-mail: [email protected]
Received Date: March 12, 2014; Accepted Date: July 18, 2014,; Published Date: July 21, 2014
Citation: Chen D, Sun Y, Zhao L, Qian Y, Zhu R, et al. (2014) Development of Two Multiplex Real-Time PCR Assays for the Rapid Detection of RNA and DNA Viruses Associated with Gastroenteritis in Pediatric Patients. Pediat Therapeut 4:208. doi:10.4172/2161-0665.1000208
Copyright: © 2014 Chen D, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Background: Viral gastroenteritis is one of the most common causes of morbidity and mortality in infants and young children in China.
Objectives: To develop a rapid and sensitive real time PCR method capable of detecting several viruses associated with gastroenteritis in pediatric patients simultaneously.
Methods: Two multiplex real-time PCR assays, the first targeting the RNA viruses: rotavirus, norovirus and parachovirus and the second the DNA viruses: human adenovirus and human bocavirus 2, were designed and assessed for their specificity and sensitivity. The multiplex assays were evaluated using clinical samples and compared to conventional RT-PCR/PCR. Results The multiplex assays developed in the study were successful in detecting the five target viruses. No cross-reactions with a panel of other human viruses were presented. The assays were sensitive enough to detect as little as one copy of in vitro transcribed target RNA or plasmid DNA in a single reaction. Compared to conventional RT-PCR or PCR, the multiplex assays showed acceptable sensitivity from 86.2% to 95.7% together with high specificity (97.2% to 99.5%) in detecting rotavirus, norovirus and adenovirus.
Conclusion: The development of these two multiplex assays should result in significant improvement in the screening of viral pathogens associated with pediatric gastroenteritis in China.