소 브루셀라병 진단을 위한 라텍스 응집반응 시험법

• 영문명: Modification and optimization of the latex agglutination test(LAT) for diagnosis of bovine brucellosis
• 발행기관: 한국예방수의학회
• 저자명: 백병걸(Byeong Kirl Baek) 허진(Jin Hur) 진찬문(Chan Moon Jin)
• 간행물 정보: 『Korean Journal of Veterinary Public Health』Vol.23, No.3, 153~163쪽, 전체 11쪽
• 주제분류: 의약학 > 면역학
• 파일형태: PDF
• 발행일자: 1999.09.30

국문 초록

브루셀라병 검진에 있어 민감도와 특이도가 높고 진단 술식이 간단한 라텍스 beads를 이용한 응집반응을 microplate에서 수행하기 위한 항원농도, 완충액의 적정조건등을 확립하기 위하여 연구하였던 바, 다음과 같은 결과를 얻었다. 1. 감작완충액으로는 0.1M TrisㆍHCI, pH 8.0을, 라텍스 beads에 감작된 항원을 안정시킬 차단완충액으로는 0.5% BSA을 함유한 0.1M TrisㆍHCI, pH 8.0을, 그리고 혈청희석완충액으로는 300mM NaCl과 0.5% BSA 및 0.01% Tween 20을 함유한 0.1M TrisㆍHCI, pH 7.4를 사용하였다. 2. 라텍스 beads에 항원을 감작시킬 감작 조건은 37℃, 60분이었으며, 라텍스 beads와 항원의 최적 농도는 모두 2.0%였다. 3. Polystyrene 라텍스 beads 부유액을 감작완충액으로 원심 세척한 다음, 농도를 2.0%로 조정한 후, PBS(pH 7.2)로 회석된 항원(2.0%)과 라텍스 beads의 배합비율이 24:1이 되도록 배합한 다음, 37℃에서 60분간 감작시켰다. 원심, 침전된 감작 항원을 차단완충액을 가하여 BWC-latex로 준비하여 4℃에 보관, 사용이 가능함을 알 수 있었다. 4. BWC-latex를 이용하여 130여두의 소 혈청에 대하여 LAT 항체 1:2 이하를 음성으로, 1:4 이상은 양성으로 하여 얻은 결과를 TAT 그리고 CFT와의 결과를 비교하였던 바, TAT 진단결과와 LAT의 결과와의 일치율에서는 민감도는 75.3%, 특이도는 100%였으며, CFT 진단결과와 LAT의 결과와의 일치율에서는 민감도와 특이도 모두 100%로 일치하였다.

영문 초록

Optimization of the latex agglutination test(LAT) for the diagnosis of bovine brucellosis was achieved by modificaion of the buffers (coating, blocking and dilution buffers) and determining the optimal incubation time and conditions of the test such as pH, ionic strength and blocking time. The optimal coating buffer was found to be 0.1M TrisㆍHCI, pH 8.0, the blocking buffer was 0.1M TrisㆍHCI, pH 8.0 containing 0.5% bovine serum albumin(BSA) and the serum dilution buffer was 0.1M TrisㆍHCI, pH 7.4 containing 300mM NaCl, 0.5% BAS and 0.01% Tween-20. One volume of a 2% latex beads suspension in coating buffer was mixed with 24 volumes of a 2% of Brucella whole cell antigen(BWC). Optimal coating period was 60min at 37℃, The resulting antigen-coated latex beads were washed two times with blocking buffer, resuspended in the same volume of the blocking buffer, called as BWC-latex. All reagents and sera were equilibrated to room temperature prior to testing and all tests were performed at ambient room temperature. Two-fold serial serum dilutions were made starting at 1:2 in 25 microliters of dilution buffer. And then the same volume of BWC-latex was added to the dilutions. The test was read after 60 min incubation at room temperature. The cut-off point for a positive LAT was established to be 1:4. Thus any sample reactive at 1:4 was considered positive. The results of the LAT were compared with the other commonly used tests such as the tube agglutination test(TAT) and the complement fixation test(CFT). In contrast to 75.3% sensitivity obtained with the TAT using 130 bovine sera, the modified LAT gave 100% specificity and sensitivity, and compared favorably with the CFT. Based on data obtained on 130 bovine serum samples, with appropriate quality control, it is concluded that the LAT is more stable and sensitive than TAT and more simple CFT. It is suggested that the LAT should be considered as a candidate for routine testing for brucellosis in Korea.

목차

ABSTRACT

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재료 및 방법

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국문요약

참고문헌

키워드

Brucellosis Tube agglutination test Complement fixation test Latex agglutination test