Abstract
The polymerase chain reaction (PCR) has become a powerful molecular diagnostic technique over the past few decades, but remains somewhat impaired due to low specificity, poor sensitivity, and false positive results. Metal and carbon nanomaterials, quantum dots, and metal oxides, can improve the quality and productivity of PCR assays. Here, we describe the ability of PCR assisted with nanomaterials (nano-PCR) comprising a nanocomposite of graphene oxide (GO) and gold nanoparticles (AuNPs) for sensitive detection of the foot-and-mouth disease virus (FMDV). Graphene oxide and AuNPs have been widely applied as biomedical materials for diagnosis, therapy, and drug delivery due to their unique chemical and physical properties. Foot-and-mouth disease (FMD) is highly contagious and fatal for cloven-hoofed animals including pigs, and it can thus seriously damage the swine industry. Therefore, a highly sensitive, specific, and practical method is needed to detect FMDV. The detection limit of real-time PCR improved by ~1000 fold when assisted by GO-AuNPs. We also designed a system of detecting serotypes in a single assay based on melting temperatures. Our sensitive and specific nano-PCR system can be applied to diagnose early FMDV infection, and thus may prove to be useful for clinical and biomedical applications.
Highlights
Foot-and-mouth disease (FMD) is a highly contagious and fatal disease of wild and domestic cloven-hoofed animals such as cattle, sheep, goat, and swine. It is caused by infection with the foot-and-mouth disease virus (FMDV)
We developed a nano-polymerase chain reaction (PCR) system for the sensitive and specific detection of FMDV by enhancing PCR with graphene oxide-gold nanoparticles nanocomposite (GO-AuNPs) that supported DNA amplification and improved PCR efficiency by interacting with PCR reagents
Serotype O- and A-type FMDV genes that originated from O-type (O/Andong/KOR/2010) and A-type (A/Pocheon/001/KOR/2010) strains were designed based on part of the RNA-dependent RNA polymerase sequence
Summary
Foot-and-mouth disease (FMD) is a highly contagious and fatal disease of wild and domestic cloven-hoofed animals such as cattle, sheep, goat, and swine. Nanomaterials, including gold and magnetic nanoparticles, titanium oxide, quantum dots, and carbon-based materials have been used to develop PCR assays with outstanding sensitivity, specificity, and efficiency. These nanomaterials are thought to improve PCR efficiency and specificity via increased thermal conductivity and interaction with PCR reagents [20,21,22,23]. We developed a nano-PCR system for the sensitive and specific detection of FMDV by enhancing PCR with graphene oxide-gold nanoparticles nanocomposite (GO-AuNPs) that supported DNA amplification and improved PCR efficiency by interacting with PCR reagents. This remarkable amplification ability would be useful for many critical circumstances, such as diagnosis of the early stages of various infectious diseases
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