Combination of polymerase chain reaction and gene chip for the rapid detection of canine viral disease

• Main Authors: Hsiu-Chiung Megan Chen, 陳秀瓊
• Other Authors: Ling-Ling Chueh
• Format: Others
• Language: zh-TW
• Published: 2002
• Online Access: http://ndltd.ncl.edu.tw/handle/43596179337691475803

碩士 === 國立臺灣大學 === 獸醫學研究所 === 90 === Rabies virus (RV), canine distemper virus (CDV), canine adenovirus (CAV) and canine parvovirus (CPV) are very important viral pathogens of dogs. Of all the four diseases, rabies belongs to zoonosis. Taiwan is at present free of this disease. However, rabies has been identified in many of the Asian countries including our neighbor mainland China. We are right now under the great risk of the outbreak of this disease. To be aware of the situation and be able to identify the disease once it is introduced, a well established diagnosis method is necessary. Moreover, the differential diagnosis of this four diseases is necessary . In this study, we perform the multiplex RT-PCR to detect and distinguish the four viruses. The PCR product was further used to hybridize specific probes by gene chip to confirm the origin of viruse e.g. field or vaccine . The results showed that the four pairs of primers designed in this study could detect RV, CDV, CAV, and CPV respectively. Moreover, all the viruses also could be detected and distinguished from each other successfully by multiplex RT- PCR. In combination with gene chip, RV, CDV, CAV, and CPV can be detected with their specific probes. Differentiation between vaccine and field strain viruses in the identification of CDV and CPV was successful. In the sensitivity assay, combination with gene chip was 100-fold more sensitive than gel electrophoresis. In the conclusion, the detection system developed in this study is rapid , sensitive and accurate method suitable for the diagnosis of clinical specimens.