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몽골 유래 Brucella melitensis 동정 및 특이 SNP를 이용한 real-time PCR법에 의한 진단 평가
강성일,김지연,김숙미,이진주,성소라,김연희,정석찬,허문,Kang, Sung-Il,Kim, Ji-Yeon,Kim, Suk Mi,Lee, Jin Ju,Sung, So-Ra,Kim, Yeon-Hee,Jung, Suk Chan,Her, Moon 대한수의학회 2015 大韓獸醫學會誌 Vol.55 No.2
A real-time PCR assay using hybridization probe (HybProbe) has been developed to detect Brucella (B.) melitensis strains. The primer and HybProbe sets were designed based on the gap gene of chromosome I with a specific single nucleotide polymorphism of B. melitensis. Specificity of the assay was confirmed by comparison to reference Brucella species and other related strains. In the melting curve analysis, B. melitensis generated a peak at $67^{\circ}C$ unlike those for other Brucella species observed at $61^{\circ}C$. Sensitivity of the assay for B. melitensis ranged from 20 ng to 200 fg of genomic DNA. The ability to identify 94 Mongolian B. melitensis isolates using the real-time PCR assay was identical to that of classical biotyping methods and differential multiplex PCR. These data showed that this new molecular technique is a simple and quick method for detecting B. melitensis, which will be important for the control and prevention of brucellosis.
성소라 ( So Ra Sung ),김지연 ( Ji Yeon Kim ),허문 ( Moon Her ),이기찬 ( Ki Chan Lee ),구정희 ( Jeong Hui Gu ),강성일 ( Sung Il Kang ),이향근 ( Hyang Keun Lee ),김숙미 ( Suk Mi Kim ),정석찬 ( Suk Chan Jung ) 한국동물위생학회 2012 韓國家畜衛生學會誌 Vol.35 No.4
A confirmatory serological test, the standard tube agglutination test (STAT) is evaluated for the diag-nostic efficiency in brucellosis Korea. A total of 345 bovine samples were collected from regional vet-erinary branch under national brucellosis monitoring program from January 2010 to June 2012 in Korea. These samples were diagnosed as suspected serum and brucellosis positive by the Rose Bengal test (RBT) and the STAT, respectively. The STAT was compared and evaluated with three serological test such as the indirect-enzyme linked immunosorbent assay (I-ELISA), competitive-enzyme linked im-munosorbent assay (C-ELISA) and fluorescence polarisation assay (FPA) prescribed for international trade by OIE. Among the 345 bovine serum samples, 302 (87.5%) were diagnosed as positive in the STAT, while 215 (62.3%), 223 (64.6%) and 194 (56.2%) serum samples were diagnosed as positive for brucellosis in the I-ELISA, C-ELISA and FPA, respectively. The STAT showed quite high positive results as compared with three prescribed tests of OIE. FPA, I-ELISA and C-ELISA have shown 60.6%, 64.9% and 67.2% correlation, respectively as compared to the STAT. However correlations of three pre-scribed tests ranged high 84.1∼97.7%. Especially, correlation between I-ELISA and C-ELISA is quite high, 97.7%. These results suggest that the STAT has shown many false-positive reactions. Therefore, additional serological test, such as ELISAs and FPA, would be necessary to adopt as a confirmatory test in the national surveillance program of bovine brucellosis in Korea.