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Báo cáo khoa học: "Development of a Lightcycler-based reverse transcription polymerase chain reaction for the detection of foot-and-mouth disease virus"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Development of a Lightcycler-based reverse transcription polymerase chain reaction for the detection of foot-and-mouth disease virus | J. Vet. Set. 2005 6 3 207-212 JOURNAL OF Veterinary Science Development of a Lightcycler-based reverse transcription polymerase chain reaction for the detection of foot-and-mouth disease virus Jae Ku Oem Soo Jeong Kye1 Kwang Nyeong Lee1 Yong Joo Kim1 Jee Yong Park1 Jong Hyeon Park1 ViS kJo liXjongSon7 National Veterinary Research and Quarantine Service Ministry of Agriculture and Forestry Anyang 430-824 Korea Department of Veterinary Infectious Disease College of Veterinary Medicine Chonbuk National University Jeonju 561-756 Korea One step TaqMan real-time reverse transcription polymerase chain reaction R T RT-PCR using a set of primers probes was developed for the detection of foot-and-mouth disease FMD virus. The gene-specific probes labeled fluorogen for the internal ribosomal entry site Leader sequence and 2B regions were used to detect FMD virus FMDV . This assay specifically detected FMDV both in cell culture preparations and clinical samples and was capable of distinguishing FMD from other viral diseases similar to clinical signs swine vesicular disease vesicular stomatitis and bovine viral diarrhea . This assay was shown to be 1000-fold more sensitive than the conventional RT-PCR method. The detection limits of this assay was 1 TCIDj ml of the FMDV RNA concentration. Quantification was obtained by a standard curves plotting threshold cycle values versus known infectivity titer. The assay was sensitive specific and rapid enough to detect FMDV RNA genome in probang samples. As such the described method is reliable and provides faster disease diagnostics than the conventional RT-PCR procedure to detect FMDV. Key words FMDV quantification TaqMan R T RT-PCR Introduction Foot and mouth disease virus FMDV is the causative agent of an economically important viral animal disease reported all over the world. Although mortality associated with foot and mouth disease FMD is usually low the disease decreases livestock productivity and for the alfected country severe .