Development of a Lightcycler-based reverse transcription polymerase chain reaction for the detection of foot-and-mouth disease virus.
- Author:
Jae Ku OEM
1
;
Soo Jeong KYE
;
Kwang Nyeong LEE
;
Yong Joo KIM
;
Jee Yong PARK
;
Jong Hyeon PARK
;
Yi Seok JOO
;
Hee Jong SONG
Author Information
1. National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry, Anyang 430-824, Korea. jku0622@nvrqs. go. kr.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
FMDV;
quantification;
TaqMan R/T RT-PCR
- MeSH:
Animals;
Foot-and-Mouth Disease/*diagnosis/virology;
Foot-and-Mouth Disease Virus/*isolation&purification;
Reverse Transcriptase Polymerase Chain Reaction/*methods;
Sensitivity and Specificity;
Taq Polymerase
- From:Journal of Veterinary Science
2005;6(3):207-212
- CountryRepublic of Korea
- Language:English
-
Abstract:
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 TCID 50 /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.
