A New High-Speed Droplet-Real-Time Polymerase Chain Reaction Method Can Detect Bovine Respiratory Syncytial Virus in Less than 10 Min

Abstract

The polymerase chain reaction (PCR) has been widely used for diagnosis of infectious diseases of domestic animals. Rapid detection of respiratory pathogens of cattle is useful for making therapeutic decisions. Therefore, we developed a new genetic-based method called droplet-real-time PCR, which can detect bovine respiratory syncytial virus (BRSV) within 10 min. Our droplet-real-time PCR markedly reduced the reaction time of reverse transcription-PCR while maintaining the same sensitivity as conventional real-time PCR, and it can be used as a rapid assay for detection of BRSV. Furthermore, our method is potentially applicable for rapid diagnosis of almost all infectious diseases, including highly pathogenic avian influenza virus.

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“…Recently, we developed novel rapid assays using a realtime droplet-polymerase chain reaction (PCR) machine (Seiko Epson Corp. Head Office, Suwa, Japan) for the detection of the human influenza virus (10,11) and the promyelocytic leukemia (PML)-retinoic acid receptor α (RARA) fusion gene (12,13), for the genotyping of single nucleotide polymorphisms (14,15) and for the detection of bovine respiratory syncytial virus (16) and breakpoint cluster region (BCR)-Abelson murine leukemia viral oncogene (ABL) fusion mRNA (17). All of these assays provide a much shorter TAT than the conventional assays currently used in clinical settings while demonstrating almost the same reactivity.…”