Pediatrics & Therapeutics
Open Access

Abstract

Development of Two Multiplex Real-Time PCR Assays for the Rapid Detection of RNA and DNA Viruses Associated with Gastroenteritis in Pediatric Patients

Dongmei Chen, Yu Sun, Linqing Zhao, Yuan Qian, Runan Zhu, Liying Liu, Liping Jia, Huijin Dong, Jie Deng and Li Deng

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.