A sensitive duplex nanoparticle-assisted PCR assay for identifying porcine epidemic diarrhea virus and porcine transmissible gastroenteritis virus from clinical specimens

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• 作者：Yu Zhu ; Lin Liang ; Yakun Luo ; Guihua Wang ; Chunren Wang ; Yudong Cui ; Xia Ai…
• 关键词：Duplex nanoPCR ; N gene ; PEDV ; TGEV ; Clinical diagnosis
• 刊名：Virus Genes
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：53
• 期：1
• 页码：71-76
• 全文大小：
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Medical Microbiology; Virology; Plant Sciences;
• 出版者：Springer US
• ISSN：1572-994X
• 卷排序：53

文摘

In this study, a novel duplex nanoparticle-assisted polymerase chain reaction (nanoPCR) assay was developed to detect porcine epidemic diarrhea virus (PEDV) and porcine transmissible gastroenteritis virus (TGEV). Two pairs of primers were designed based on the conserved region within the N gene of PEDV and TGEV. In a screening of 114 clinical samples from four provinces in China for PEDV and TGEV, 48.2 and 3.5 % of the samples, respectively, tested positive. Under optimized conditions, the duplex nanoPCR assay had a detection limit of 7.6 × 101 and 8.5 × 101 copies μL−1 for PEDV and TGEV, respectively. The sensitivity of the duplex nanoPCR assay was ten times higher than that of a conventional PCR assay. Moreover, no fragments were amplified when the duplex nanoPCR assay was used to test samples containing other porcine viruses. Our results indicate that the duplex nanoPCR assay described here is useful for the rapid detection of PEDV and TGEV and can be applied in clinical diagnosis.