| Isolation of akabane virus and its molecular diagnosis by reverse transcription polymerase chain reaction |
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Jae-jin Cho1, Chung-gil Lee2, Bong-kyun Park3, Chung-ho Chang1, Chung-won Chung1, In-soo Cho1, Soo-hwan An1 |
1National Veterinary Research and Quarantine Service
2College of Veterinary Medicine, Chonnam National University
3College of Veterinary Medicine, Seoul National University |
| 아까바네 바이러스의 분리 및 RT-PCR 진단법에 관한 연구 |
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조재진1, 이정길2, 박봉균3, 장정호1, 정정원1, 조인수1, 안수환1 |
1국립수의과학검역원
2전남대학교 수의과대학
3서울대학교 수의과대학 |
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| Abstract |
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Akabane disease is transmitted through mosquitoes in cattle, sheep and goats. It shows congenital abnormalities including encephalomyetitis, hydranencephaly, neurogenic arthrogryposis, and deformed neonatal calves. Akabane viruses, 93FMX and K-9 strain, were isolated from fetal matrix of aborted cow and blood of healthy cow, respectively. S gene sequences of 93FMX and K-9 showed 100% homology with that of OBE-1 strain isolated in Japan. Based upon our sequencing data, we synthesized specific primers for PCR diagnosis. Using these primers, we were able to amplify the S gene of Akabane virus not only from the culture fluid of Vero cells but also from the brain tissue of suckling mouse inoculated with, Akabane virus. These PCR products were confirmed by Southern blot hybridization. Not only the sensitivity of PCR test was high enough to detect the viruses of $10^{1.0}TCID_{50}/ml$, but also the time for diagnosis was significantly shorter than that of the virus isolation by tissue culture method. This method was also effective for the detection of Akabane virus in the cerebrum of fetus. RT-PCR method may be used for a useful diagnostic test of the clinical cases of Akabane disease. |
| Key Words:
Akabane virus, PCR, virus isolation |
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