국내 분리 돼지 유행성설사 바이러스 Chinju99주의 생물학적 및 물리화학적 성상 Biological and Physicochemical Properties of Porcine Epidemic Diarrhea Virus Chinju99 Strain Isolated in Korea원문보기
진주 지역에서 1000 여두의 생후 1주령 포유 자돈군에 심한 설사증이 발생하였던바 소장조직으로부터 RNA를 추출한 후 reverse transcription-polymerase chain reaction을 실시하였던 결과, porcine epidemicdiarrhea (PED) virus (PEDV)의 N 유전자가 검출되어 PED로 진단되었다. Trypsin (10ug/ml)을 첨가한 배지에서 Vero 세포를 배양하면서 장조직으로부터 바이러스 분리배양을 시도하였던 결과 2회의 blind passage 후에 PEDV를 분리할 수 있었다. 따라서 이 분리주를 Chinju99로 명명하였으며, Chinju99주의 생물학적 및 물리화학적 성상을 조사하였던 결론은 다음과 같다. Chinju99주는 전자현미경 소견에서 비정형의 타원형 입자로서 표면에 spike 구조를 가지고 있었으며, Vero 세포내 배양시에 점진적으로 원형변성, 합포체형성 등의 세포변성 소견을 나타내었다. 또한 20% ether, 5% chloroform에서 불안정하였으며 pH 4-7의 산성조건에서는 안정성을 나타내었다. 동시에 $50^{\circ}C$에서 180분간 처리 후에도 감염성이 유지되었고 sucrose 용액에서의 부유밀도가 1.180g/ml으로서, coronavirus의 전형적인 생물학적 및 물리화학적 성상을 나타내었다.
진주 지역에서 1000 여두의 생후 1주령 포유 자돈군에 심한 설사증이 발생하였던바 소장조직으로부터 RNA를 추출한 후 reverse transcription-polymerase chain reaction을 실시하였던 결과, porcine epidemic diarrhea (PED) virus (PEDV)의 N 유전자가 검출되어 PED로 진단되었다. Trypsin (10ug/ml)을 첨가한 배지에서 Vero 세포를 배양하면서 장조직으로부터 바이러스 분리배양을 시도하였던 결과 2회의 blind passage 후에 PEDV를 분리할 수 있었다. 따라서 이 분리주를 Chinju99로 명명하였으며, Chinju99주의 생물학적 및 물리화학적 성상을 조사하였던 결론은 다음과 같다. Chinju99주는 전자현미경 소견에서 비정형의 타원형 입자로서 표면에 spike 구조를 가지고 있었으며, Vero 세포내 배양시에 점진적으로 원형변성, 합포체형성 등의 세포변성 소견을 나타내었다. 또한 20% ether, 5% chloroform에서 불안정하였으며 pH 4-7의 산성조건에서는 안정성을 나타내었다. 동시에 $50^{\circ}C$에서 180분간 처리 후에도 감염성이 유지되었고 sucrose 용액에서의 부유밀도가 1.180g/ml으로서, coronavirus의 전형적인 생물학적 및 물리화학적 성상을 나타내었다.
A disease with severe diarrhea occurred in a herd of one thousand, 1-week-old piglets in Chinju, Korea, and was diagnosed as porcine epidemic diarrhea by the detection of N gene of porcine epidemic diarrhea virus (PEDV) from small intestines. A PEDV, named as Chinju99, was also isolated from the int...
A disease with severe diarrhea occurred in a herd of one thousand, 1-week-old piglets in Chinju, Korea, and was diagnosed as porcine epidemic diarrhea by the detection of N gene of porcine epidemic diarrhea virus (PEDV) from small intestines. A PEDV, named as Chinju99, was also isolated from the intestines after two blind-passages in Vero cells supplemented with trypsin (10 ug/ml). and the biological and physicochemical properties of the isolate were characterized. The virion was roughly spherical in shape and had spike peplomers on its outer surface. The virus exhibited cytopathic effects such as rounding degeneration at initiation of infection and syncytia formation later in Vero cells. The virus was labile to 20% ether and 5% chloroform but stable in acid with pH 4-7 at $4^{\circ}C$. The infectivity of the virus was maintained at $50^{\circ}C$ for 180 min, and the buoyant density of the virus in sucrose was 1.180 g/ml. All biological and physicochemical properties of the virus were typical features of coronaviruses.
A disease with severe diarrhea occurred in a herd of one thousand, 1-week-old piglets in Chinju, Korea, and was diagnosed as porcine epidemic diarrhea by the detection of N gene of porcine epidemic diarrhea virus (PEDV) from small intestines. A PEDV, named as Chinju99, was also isolated from the intestines after two blind-passages in Vero cells supplemented with trypsin (10 ug/ml). and the biological and physicochemical properties of the isolate were characterized. The virion was roughly spherical in shape and had spike peplomers on its outer surface. The virus exhibited cytopathic effects such as rounding degeneration at initiation of infection and syncytia formation later in Vero cells. The virus was labile to 20% ether and 5% chloroform but stable in acid with pH 4-7 at $4^{\circ}C$. The infectivity of the virus was maintained at $50^{\circ}C$ for 180 min, and the buoyant density of the virus in sucrose was 1.180 g/ml. All biological and physicochemical properties of the virus were typical features of coronaviruses.
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제안 방법
Following the method of Hofmann and Wyler8, the virus of 104 PFU/ml was diluted to 1:10 in MEM-T and pH of the viral fluid dilutions was adjusted to 2, 3, 4, 5, 6, and 7, respectively, with 2N HCL After acidic treatment at 4℃ for 6 h, pH of the viral fluid was readjusted to 7.4 with 2N NaOH. Propagation ability of the virus in Vero cells cultured with MEM-T was observed by plaque assay.
The piglets suffered from diarrhea, vomiting and dehydration, and the disease was presumptively diagnosed as PED based on the clinical signs. Intestinal tissues and feces of these piglets were collected and transported to the laboratory The samples were emulsified with 10 volumes of minim시 essential medium (MEM) (Invitrogen, USA) and used for the detection of viral gene and isolation of PEDV. The tissue emulsion, which was used for the viral isolation, was created with streptomycin (500 |ig/ml) and penicillin (500 U/ml) at 37℃ for 1 h, and clarified by centrifugation at 5000 >< g for 30 min.
성능/효과
In conclusion, all biological and physicochemical properties of the PEDV Chinju99 was characteristic features of coronavirusese13-14. To our knowledge, the biological and physicochemical properties of a PEDV isolated in Korea were first reported in the present study.
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