Comparison of fluorescent and conventional darkfield methods for the detection of Treponema pallidum in syphilitic lesions.
Comparison of fluorescent and conventional darkfield methods for the detection of Treponema pallidum in syphilitic lesions.
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荧光和传统暗场方法检测梅毒病变中梅毒螺旋体的比较。
DOI：
10.1093/ajcp/47.6_ts.809
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发表时间：
1967
期刊：
影响因子：
--
通讯作者：
W. Ketterer
中科院分区：
文献类型：
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作者：
R. Jue;J. Puffer;R. Wood;G. Schochet;W. H. Smartt;W. Ketterer
在这篇文章中，作者比较了使用荧光抗体和传统暗地方法检测梅毒螺旋体的结果。对于荧光检测 (FADF)，梅毒血清的球蛋白部分通过硫酸铵分离，并以 40:1 的蛋白质-染料比例与异硫氰酸荧光素缀合；因为这种结合物与其他螺毛菌一起发出明亮的荧光，所以它被雷特密螺旋体吸收，过程有些复杂（详细信息应查阅原文）；测试表明，吸收后，缀合物仅对T. 的致病菌株进行染色。苍白球。对于荧光测试，将缀合物用等量的吐温 80 在磷酸盐缓冲盐水 (pH 7.2) 和无菌正常人血清中稀释 1/10；然后将其涂在待测涂片上，并在 37°C 的潮湿室中孵育 30 分钟；将载玻片先用缓冲盐水漂洗，然后用蒸馏水漂洗，封固并在荧光显微镜下以 400 倍放大倍数进行检查。所检测的 241 个病变中有 58 个被证实存在苍白球，其中 52 个荧光呈阳性，44 个暗底呈阳性； 38例两者均呈阳性； 20 个病变给出了不同的结果； 14 例 FADF 呈阳性，6 例仅通过暗底检测呈阳性。讨论差异； FADF 似乎比暗地更敏感，但至少给出了一个假阳性结果。据称 FADF 与暗地一样准确，并且不依赖于运动密螺旋体的存在 [但肯定这些生物体的运动是其最典型的特征]。 E.奥斯蒙德.
In this article the authors compare results of tests forTreponema pallidumusing fluorescent antibody and conventional dark ground methods.For the fluorescence test (FADF) the globulin fraction of syphilitic serum was separated by ammonium sulfate and conjugated with fluorescein isothio-cyanate with a 40: 1 protein-dye ratio; because this conjugate gave bright fluorescence with other spiro-chaetes it was absorbed with Reiter treponemes by a somewhat complicated process (for details the original should be consulted); tests showed that after absorption the conjugate stained only the pathogenic strain ofT. pallidum. For the fluorescence test the conjugate was diluted 1/10 with equal parts of Tween 80 in phosphate buffered saline (pH 7.2) and sterile normal human serum; this was then applied to smears to be tested which were incubated in a moist chamber at 37°C for 30 minutes; the slides were rinsed in buffered saline and then distilled water, mounted and examined at X 400 magnification under a fluorescence microscope.T. pallidumwas demonstrated in 58 of the 241 lesions tested, 52 being positive by fluorescence and 44 by dark ground; both were positive in 38 cases; 20 lesions gave discrepant results; 14 were positive with FADF and 6 by dark ground only. The discrepancies are discussed; it seems that FADF is rather more sensitive than dark ground but gave at least one false positive result.It is claimed that FADF is as accurate as dark ground and does not depend on the presence of motile treponemes [but surely the movement of these organisms is their most characteristic feature].T. E. Osmond.