The Korean Journal of Medicine

The Korean Association of Internal Medicine (대한내과학회)
권호 표지

Identification of Brucella abortus using the sequencing of omp gene

주외막단백 유전자 분석을 이용한 Brucella abortus의 동정

Lee, Yu-Ji;Ko, Kwan-Soo;Park, Mi-Yeoun;Oh, Won-Sup;Kwon, Ki-Tae;Ryu, Seong-Yeol;Heo, Sang-Taek;Peck, Kyong-Ran;Lee, Nam-Yong;Song, Jae-Hoon
이유지;고관수;박미연;오원섭;권기태;류성열;허상택;백경란;이남용;송재훈

  • Published : 20060700
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Abstract

Background : As the incidence of bovine brucellosis increases in Korea, the incidence of human brucellosis is also increasing since 2002. However, it is difficult to identify Brucella species by using the conventional methods. Methods : Three strains of gram-negative coccobacilli were isolated from blood specimens of three patients with prolonged fever, which were not identified by using the conventional methods. After extracting total DNA from these isolates, PCR amplification of 16S rRNA and omp2 genes was performed. These sequences secured by PCR assay were compared with known sequences by using GenBank BLAST. Results : DNA sequences were obtained from 3 isolates by using PCR amplification of 16S rRNA. These sequences had more than 99.9% similarities with Brucella species by using GenBank BLAST. In the second place, after comparing DNA sequences secured by PCR amplification of omp2a and omp2b by using GenBank BLAST, these isolates were confirmed as B. abortus. Conclusions : DNA sequence analysis is a rapid and accurate method for identification of uncommon microorganisms, such as Brucella species.

목적 : 브루셀라증은 다양한 임상경과와 비특이적인 증상 및 까다로운 배양 조건 등으로 인하여 진단이 쉽지 않다. 최근 국내에서 발생한 발열 환자의 혈액 배양검사로부터 균주를 분리한 후 유전자 염기서열분석으로 Brucella abortus를 확인하였기에 이를 소개하고자 하였다. 방법 : 발열 환자의 혈액배양검사로부터 분리된 미상의 균주로부터 DNA를 추출한 후 16S rRNA에 대한 연쇄중합반응을 시행하였고, 증폭된 결과물을 BLAST search를 통하여 GenBank의 데이터베이스의 염기서열과 비교한 결과 Brucella종임을 확인하였다. 동일한 방법으로 omp2a와 omp2b 유전자에 대한 연쇄중합반응을 시행하여 이를 GenBank의 데이터베이스와 비교하였다. 결과 : 16S rRNA 염기서열을 비교하여 세 균주 모두 99.9% 이상의 유사도로 Brucella에 속하는 종임을 확인하였고, omp2a와 omp2b 유전자 염기서열을 비교한 결과 99.91% 이상의 유사도로 세 균주 모두 Brucella abortus임을 확인하였다. 결론 : 유전자 염기서열분석은 비교적 짧은 시간 내에 정확한 균종의 확인이 가능하다는 이점이 있어서 생물형 분석으로 동정되지 않는 드문 균주의 동정에 매우 유용한 방법으로 생각된다.

Keywords

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