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墨西哥用于诊断牛布鲁氏菌病的荧光偏振检测法评估

Evaluation of a fluorescence-polarization assay for the diagnosis of bovine brucellosis in México.

作者信息

Dajer A, Luna-Martínez E, Zapata D, Villegas S, Gutiérrez E, Peña G, Gurría F, Nielsen K, Gall D

机构信息

Universidad Autónoma de Yucatán, Facultad de Medicina Veterinaria y Zootecnia, Depto. de Inmunología. km., Yucatan, Mexico.

出版信息

Prev Vet Med. 1999 May 14;40(1):67-73. doi: 10.1016/s0167-5877(99)00004-5.

Abstract

A homogeneous fluorescence-polarization assay (FPA) was used for the serological diagnosis of bovine brucellosis in México. The assay uses O-polysaccharide prepared from Brucella abortus lipoplysaccharide (20-30 kDa) conjugated with fluorescein isothiocyanate as a tracer. To measure the fluorescence polarization, a FPM-1 fluorescence-polarization analyzer was used with the procedure described by Nielsen et al. (1996b). A cut-off value of 90 millipolarization (mP) units was used for testing 560 bovine sera from different areas of México. (305 positive sera and 255 negative sera according to the complement fixation test; CFT.) Some were tested with the Rose Bengal plate (RB) test (n = 490) and some with the rivanol-agglutination (RIV) test (n = 190). Sensitivities were 98.3%, 99.3% and 99.0%, and specificities were 68.8%, 55.4% and 96.9%, respectively, for RB, RIV and FPA. The FPA gave a kappa coefficient of agreement with respect to CFT of 0.96, while RB and RIV (relative to the CFT) gave coefficients of 0.70 and 0.61, respectively. Finally, ROC analysis suggested a cut-off value which agreed with the one recommended in the test procedure. We concluded that FPA is a suitable test to be used instead of the CFT in Mexican conditions.

摘要

一种均相荧光偏振分析方法(FPA)被用于墨西哥牛布鲁氏菌病的血清学诊断。该分析方法使用从流产布鲁氏菌脂多糖(20 - 30 kDa)制备的O - 多糖与异硫氰酸荧光素偶联作为示踪剂。为测量荧光偏振,使用了FPM - 1荧光偏振分析仪,采用Nielsen等人(1996b)描述的程序。使用90毫偏振(mP)单位的临界值对来自墨西哥不同地区的560份牛血清进行检测。(根据补体结合试验;CFT,其中305份为阳性血清,255份为阴性血清。)一些血清用虎红平板(RB)试验检测(n = 490),一些用利凡诺凝集(RIV)试验检测(n = 190)。RB、RIV和FPA的灵敏度分别为98.3%、99.3%和99.0%,特异性分别为68.8%、55.4%和96.9%。FPA相对于CFT的kappa一致性系数为0.96,而RB和RIV(相对于CFT)的系数分别为0.70和0.61。最后,ROC分析得出的临界值与检测程序中推荐的一致。我们得出结论,在墨西哥的条件下,FPA是一种适合替代CFT的检测方法。

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