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Title: COMPARISON BETWEEN REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION (RT-PCR) AND REAL-TIME RT-PCR FOR DETECTION OF FOOT-AND-MOUTH DISEASE VIRUS (FMDV)

Author
item KONIG, GUIDO - INTA, CASTELAR ARGENTIA
item Piccone, Maria
item CARRILLO, ELISA - INTA, CASTELAR ARGENTINA

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 9/1/2005
Publication Date: 9/19/2005
Citation: Konig, G.A., Piccone, M.E., Carrillo, E. 2005. Comparison between reverse transcription polymerase chain reaction (rt-pcr) and real-time rt-pcr for detection of foot-and-mouth disease virus (fmdv). Meeting Abstract. VIII Congreso Argentino de Virologia. Page 37, Number 10561.

Interpretive Summary:

Technical Abstract: As part of Argentina's national preparedness for the rapid detection and diagnosis of foot-and-mouth disease (FMD) outbreakes, we have developed a reverse transcription polymerase chain reaction (RT-PCR) for the detection of FMDV. This assay that targeted the viral 3D polymerase-coding region specifically detected FMDV and was capable of distinguishing FMD from other viral diseases with similar clinical signs. The detection limit of the assay was 7 pg of vARN or 1600 PFU. Currently, the laboratory of the National Animal Health Service (SENASA, Argentina) runs this RT-PCR on highly suspect FMD samples by EITB assay or antigen captures ELISA test used for diagnosis of FMDV. In this work we described the development of a real-time reverse transcription polymerase chain reaction and compared it to the conventional RT-PCR. We used the same set of primers (3D region) and the Sybr Green kit (Applied Biosystems). The following parameters were optimized: cycling number and profile, Mg++ and NTPs concentration. This assay was shown to be more sensitive than the convetional RT-PCR method. In conclusion; this study demonstrated that the real-time RT-PCR provides an extremely sensitive and rapid procedure for disease diagnosis than the conventional RT-PCR.