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      • Canine Distemper Virus 검출을 위한 정량 Real-time PCR

        이승관,이창규,정운원,조경진,김현숙,강영대,서장훈 高麗大學校 倂設 保健大學 保健科學硏究所 2002 保健科學論集 Vol.28 No.2

        Canine distemper virus(CDV) infection causes a fatal systemic disease in wide range of carnivore species, including domestic dogs. It has been a highly contagious disease of veterinary importance for centurise. Recently many researches show that qualitative RT-PCR for the detection of CDV was sensitive and specific method for the early and safe antemortem diagnosis of distemper by using whole blood regardless of clinical sign and pathological finding. Recently, results were correlated to clinical findings, anti-CDV neutralizing antibody titers, postmortem findings, and demonstration of CDV NP(nucleoprotein) antigen by immunohistochemistry. We have studied a quantitative real-time PCR assay for CDV in order to improve and broaden the clinical usefulness of RT-PCR assay. Total 37 samples including 9 clinically positive and 29 randomly selected ones have investigated. CDV RNA was extracted from whole blood. The isolated RNA was reverse-transcripted into cDNA followed by PCR amplification. The assay based on real-time PCR using SYBR Green I dye as quantitative method with a LightCycler. Calibration curves covered 30-3×10^(8) copies of CDV were created using cloned 240bp CDV cDNA of viral RNA. CDV was detected all in 8 positive samples whereas 8 in 29 randomly collected ones. The detection ranges of clinical CDV specimens were various from 3×10² to 3×10⁴ copies. Comparision of real-time PCR to conventional PCR showed equal sensitivity and specificity. But real-time PCR method has a merit of quantification as well as more rapid results than those of common RT-PCR. CDV can be accurately detected and quantified in the whole blood from the dog infected with CDV. This quantitative technique may improve the clinical utility of RT-PCR for the detection of CDV.

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