Detection of Babesia canis and Babesia gibsoni by Polymerase Chain Reaction

• Main Authors: Lin, Cheng-Yu, 林政佑
• Other Authors: Andrew Chang-Young Fei
• Format: Others
• Language: zh-TW
• Published: 1998
• Online Access: http://ndltd.ncl.edu.tw/handle/86316724467513766746

碩士 === 國立臺灣大學 === 獸醫學系研究所 === 86 === This study which apply molecular biological technology - polymerase chain rea ction can detect the canine babesiosis successfully. Two primers (BS1/BS2) de s igned from the 16S ribosomal RNA gene sequences were used to amplify the ge nom ic DNA of Babesia canis and Babesia gibsoni. BS1 primer : 5'-GCA TGT CTA AGT A CA AGC TTT-3' (32-52 base), BS2 primer : BS2 : 5'-GGA TTC CCA TCA TTC C AA T-3' (469-487 base) and the PCR product was 456 bp. BS1/BS primer could d etect bot h the infections of B. canis and B. gibsoni after several tests, th erefore thi s technology can be applied for rapid diagnosis. Due to the high similarity of 16S ribosomal RNA gene of B. canis and B. gibsoni, The simple PCR didn*t diff erentiate between B. canis and B. gibsoni. This research inte nded to apply Ran dom Amplified Polymorphic DNA PCR (RAPD-PCR) to differentia te the two kinds of canine babesial parasites. We applied the twenty random primers to amplify th e genomic DNA of B. canis and B. gibsoni. The results s howed that PCR amplifie d products by thirteen primers including OPO-01, OPO- 03, OPO-04, OPO-05, OPO-0 6, OPO-07, OPO-10, OPO-11, OPO-12, OPO-13, OPO-14, OPO-15, and OPO-20 had no d ifferent patterns in electrophoresis analysis. Th e PCR amplified products by a nother six primers including OPO-02, OPO-08, OP O-09, OPO-16, OPO-18, and OPO-1 9 had obvious different ones in electrophores is analysis. There was no PCR amp lified product by OPO-17 primer. It can dif ferentiate the two kind of canine b abesial parasites definitively by RAPD-PC R. According to the results, combinat ion simple PCR with RAPD-PCR technology can detect and differentiate B. canis and B. gibsoni successfully. For the clinical diagnosis of canine babesial par asites, it set up a rapid and accur ate technology.