국내 독일 셰퍼드로부터 에를리히아증과 라임병에 대한 항체가 조사를 위하여 효소면역측정법을 시용하였으며, 중합효소연쇄반응을 이용하여 진드기매개성 병원체를 조사하였다. 2005년 10월부터 2006년 9월까지 국내 5개 지방으로부터 291마리 독일 셰퍼드를 선정하였다. 개 에를리히아 항체 조사를 위한 ELISA (Snap$^{(R)}$ 3Dx$^{(R)}$ test kit, IDEXX) 검사결과 전국적으로 7.56%(22마리)의 에를리히아 양성 항체 보균개와 1.72%(5마리)의 보렐리아 양성 항체 보균개로 조사되었다. 이 중 2마리에서는 두 가지 병원체 모두에 대한 양성 항체가 검출되었다. 에를리히아에 대한 양성항체를 지방별로 보면 경기도는 1.28%(78마리 중 1마리), 강원도는 12.64%(87마리 중 11마리), 충청도는 9.76%(41마리 중 4마리), 경상도는 8.93%(56마리 중 5마리), 전라도는 3.45%(56마리 중 5마리)로 조사되었다. Ehrlichia chaffeensis 유전자를 검출한 결과 전국 평균이 3.09%(291 마리 중 9마리), 강원도는 2.41%(291 마리 중 7마리) 그리고 충청도는 0.69%(291마리중 2마리)로 조사되었다. 중합효소연쇄반응으로 증폭된 유전자(SNU-EC3과 SNU-EC5)에 대한 유전자염기서열분석결과 한국(Haemaphysalis longicornis 진드기)과 중국 그리고 Arkansas 분리주인 E. chaffeensis와 매우 유사하였다. 이 연구의 결과 국내에서 사육되는 독일셰퍼드에서 에를리히아증의 병원체와 라임병에 대한 항체가 검출된 것으로 볼 때 임상 진단에 있어서 진드기매개전염병에 대한 중요성을 인식해야 할 것으로 판단된다. 따라서 혈소판감소증이나 백혈구감소증이 있는 경우 그리고 봄부터 가을까지 야외활동을 한 경력이 있는 환자에 대한 감별진단 항목에 반드시 포함시켜야 할 것을 제안한다.
국내 독일 셰퍼드로부터 에를리히아증과 라임병에 대한 항체가 조사를 위하여 효소면역측정법을 시용하였으며, 중합효소연쇄반응을 이용하여 진드기매개성 병원체를 조사하였다. 2005년 10월부터 2006년 9월까지 국내 5개 지방으로부터 291마리 독일 셰퍼드를 선정하였다. 개 에를리히아 항체 조사를 위한 ELISA (Snap$^{(R)}$ 3Dx$^{(R)}$ test kit, IDEXX) 검사결과 전국적으로 7.56%(22마리)의 에를리히아 양성 항체 보균개와 1.72%(5마리)의 보렐리아 양성 항체 보균개로 조사되었다. 이 중 2마리에서는 두 가지 병원체 모두에 대한 양성 항체가 검출되었다. 에를리히아에 대한 양성항체를 지방별로 보면 경기도는 1.28%(78마리 중 1마리), 강원도는 12.64%(87마리 중 11마리), 충청도는 9.76%(41마리 중 4마리), 경상도는 8.93%(56마리 중 5마리), 전라도는 3.45%(56마리 중 5마리)로 조사되었다. Ehrlichia chaffeensis 유전자를 검출한 결과 전국 평균이 3.09%(291 마리 중 9마리), 강원도는 2.41%(291 마리 중 7마리) 그리고 충청도는 0.69%(291마리중 2마리)로 조사되었다. 중합효소연쇄반응으로 증폭된 유전자(SNU-EC3과 SNU-EC5)에 대한 유전자염기서열분석결과 한국(Haemaphysalis longicornis 진드기)과 중국 그리고 Arkansas 분리주인 E. chaffeensis와 매우 유사하였다. 이 연구의 결과 국내에서 사육되는 독일셰퍼드에서 에를리히아증의 병원체와 라임병에 대한 항체가 검출된 것으로 볼 때 임상 진단에 있어서 진드기매개전염병에 대한 중요성을 인식해야 할 것으로 판단된다. 따라서 혈소판감소증이나 백혈구감소증이 있는 경우 그리고 봄부터 가을까지 야외활동을 한 경력이 있는 환자에 대한 감별진단 항목에 반드시 포함시켜야 할 것을 제안한다.
The presence of the tick-borne pathogens Ehrlichia and Borrelia in German Shepherd dogs in Korea was determined by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). A total of 291 dogs were randomly selected from five Korean provinces from October 2005 through September ...
The presence of the tick-borne pathogens Ehrlichia and Borrelia in German Shepherd dogs in Korea was determined by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). A total of 291 dogs were randomly selected from five Korean provinces from October 2005 through September 2006. The seroprevalence of antibodies to canine Ehrlichia and Borrelia agents detected by ELISA (Snap$^{(R)}$ 3Dx$^{(R)}$ Test, IDEXX Laboratories) was 7.56% (22 dogs) and 1.72% (5 dogs) respectively, throughout the country. Positive antibodies against both pathogens were detected in two dogs (0.69%). The provincial distribution of seroprevalence against Ehrlichia was 1.28% (1 of 78) in Gyeonggi-do, 12.64% (11 of 87) in Gangwon-do, 9.76% (4 of 41) in Chungcheong-do, 8.93% (5 of 56) in Gyeongsang-do, and 3.45% (1 of 29) in Jeolla-do. According to PCR analysis, Ehrlichia chaffeensis target DNA was amplified in 3.09% (9 of 291 dogs) of blood samples, 2.41% (7 of 291) from Gangwon-do and 0.69% (2 of 291) from Chungcheong-do. The oligonucleotide sequences (SNU-EC3 and SNU-EC5) from the PCR fragment examined in Korea were closely related to E. chaffeensis isolated from the tick Haemaphysalis longicornis, in China and the state of Arkansas in the US. Based on these results, the presence of E. chaffeensis infection was identified in German Shepherds being bred in Korea. These results bring to light the importance of paying close attention to tick-borne infections such as Lyme disease during clinical diagnosis. This infectious disease should be included as a differential diagnosis for patients who participate in outdoor activity from spring to fall or who have thrombocytopenia or leucopenia.
The presence of the tick-borne pathogens Ehrlichia and Borrelia in German Shepherd dogs in Korea was determined by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). A total of 291 dogs were randomly selected from five Korean provinces from October 2005 through September 2006. The seroprevalence of antibodies to canine Ehrlichia and Borrelia agents detected by ELISA (Snap$^{(R)}$ 3Dx$^{(R)}$ Test, IDEXX Laboratories) was 7.56% (22 dogs) and 1.72% (5 dogs) respectively, throughout the country. Positive antibodies against both pathogens were detected in two dogs (0.69%). The provincial distribution of seroprevalence against Ehrlichia was 1.28% (1 of 78) in Gyeonggi-do, 12.64% (11 of 87) in Gangwon-do, 9.76% (4 of 41) in Chungcheong-do, 8.93% (5 of 56) in Gyeongsang-do, and 3.45% (1 of 29) in Jeolla-do. According to PCR analysis, Ehrlichia chaffeensis target DNA was amplified in 3.09% (9 of 291 dogs) of blood samples, 2.41% (7 of 291) from Gangwon-do and 0.69% (2 of 291) from Chungcheong-do. The oligonucleotide sequences (SNU-EC3 and SNU-EC5) from the PCR fragment examined in Korea were closely related to E. chaffeensis isolated from the tick Haemaphysalis longicornis, in China and the state of Arkansas in the US. Based on these results, the presence of E. chaffeensis infection was identified in German Shepherds being bred in Korea. These results bring to light the importance of paying close attention to tick-borne infections such as Lyme disease during clinical diagnosis. This infectious disease should be included as a differential diagnosis for patients who participate in outdoor activity from spring to fall or who have thrombocytopenia or leucopenia.
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문제 정의
Few human cases of borreliosis and detection of the agent from Ixodes ticks exist in Korea. The purpose of this study was to survey the prevalence of canine Ehrlichia and Borrelia infections in outdoor German Shepherd dogs in Korea.
제안 방법
Amplified and purified DNA was prepared for direct sequencing using a QIAquick Gel Extinction Kit (QIAGEN Inc., Germany) and seuenced by dideoxy termination widi an automatic sequencer (ABI 3100 Genetic Analyzer, Bionics, Korea). The sequence data were collected using ABI Prism Data Collection software (ver.
Fig 2. Phylogenetic tree generated using maximum-likelihood estimation (PAUP* 4.0b for Macintosh™) of 16S rRNA gene sequences from 23 representative E. chaffeensis SNU-EC3 and SNUE-C5 sequences amplified from German Shepherd dogs in Gangwon-do, Korea.
Shepherd dogs in Gangwon-do, Korea. The tree was constructed after aligning the 16S rRNA gene fragment sequence (390 bp) obtained from GenBank and sequenced during this study with the ClustalX program. The scale represents a number of substitutions per base and indicates horizontal distance.
대상 데이터
Blood samples were collected from 291 dogs from five provinces, Gangwon-do (n = 87), Chungcheong-do (n = 41), Gyeo- ngsang-do (n = 56), Jeolla-do (n = 29), Seoul and Gyeonggi-do (n = 78) in Korea between October 2005 and September 2006. Dogs sampled were from annual training programs at working dog training centers.
, Germany) and seuenced by dideoxy termination widi an automatic sequencer (ABI 3100 Genetic Analyzer, Bionics, Korea). The sequence data were collected using ABI Prism Data Collection software (ver.2.1), and analyzed by ABI Prism Sequence analysis software (ver. 2.1.1) and Chromas software (ver. 1.51) (Technelysium Pty., Ltd., Queensland, Australia).
데이터처리
Aligned sequences were determined by the neighbor-joining (NJ) method with nucleotide distances (p-distance) with 100 replications by the bootstrap test. Phylogenetic analysis based on the obtained sequences was conducted using maximumlikelihood estimation (PAUP* 4.0b for Macintosh™).
이론/모형
chaffeensis were used for phylogenetic analysis. Aligned sequences were determined by the neighbor-joining (NJ) method with nucleotide distances (p-distance) with 100 replications by the bootstrap test. Phylogenetic analysis based on the obtained sequences was conducted using maximumlikelihood estimation (PAUP* 4.
0) statistical computer program. Chi-square (x2) tests and Fisher exact tests were used to compare each of the prevalence values among age, sex, and geographical distribution. P < 0.
성능/효과
In terms of sex distribution, there was no significant difference in the prevalence rate, which was 6.30% (8 of 127) in female and 8.54% (14 of 164) m male dogs based on the sero- logic ELISA test and 2.36% (3 of 127) in female and 3.66% (6 of 164) in male dogs based on the PCR analysis (Table 6). The regional distribution of seroprevalence was 1 of 78 (1.
Positive Snap€3Dx® srtologic ELISA results were identified in 14 of 130 (10.77%) 4-6 year olds and 8 of 78 (10.26%) 7-9 year olds (Table 5), while PCR analysis was positive in 3 of 130 (2.31%) 4-6 year olds and 6 of 78 (7.69%) 7-9 year olds (Table 5).
The sensitivity of this test for detecting ehrlichiosis and borreliosis is 98.9% and 95.0%, and the specificity is 98.2% and 99.9% respectively, according to information provided by the manufacturer.
chaffeensis available in the GenBank database. The sequence identity of E. chaffeensis SNU-EC3 and SNU-EC5 (present study) were 99.7% and 99.2% compared to a Korean strain from tick (AY350424), USA isolate (AF416764), and Chinese isolate (AF147752), respectively. Comparative analysis of nucleotide sequences of Korean strains determined in this study with the 16S rRNA gene fragment sequences of 22 known Ehrlichia species were available in the GenBank database.
canis within sera (42). The study revealed a seroprevalence of 53% for E. canis. At that time, E.
후속연구
This study has enabled us to provide further information on the epidemiology of tick-associated bacteria in Korea, where little information on this subject exists. Further studies are required for a detailed understanding of these newly emerging tick-bome diseases in Korea.
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