Mason D J, Shanmuganathan S, Mortimer F C, Gant V A
Infection and Immunity Laboratory, United Medical School, Guy's Hospital, London, United Kingdom.
Appl Environ Microbiol. 1998 Jul;64(7):2681-5. doi: 10.1128/AEM.64.7.2681-2685.1998.
The fluorescent nucleic acid binding dyes hexidium iodide (HI) and SYTO 13 were used in combination as a Gram stain for unfixed organisms in suspension. HI penetrated gram-positive but not gram-negative organisms, whereas SYTO 13 penetrated both. When the dyes were used together, gram-negative organisms were rendered green fluorescent by SYTO 13; conversely, gram-positive organisms were rendered red-orange fluorescent by HI, which simultaneously quenched SYTO 13 green fluorescence. The technique correctly predicted the Gram status of 45 strains of clinically relevant organisms, including several known to be gram variable. In addition, representative strains of gram-positive anaerobic organisms, normally decolorized during the traditional Gram stain procedure, were classified correctly by this method.
荧光核酸结合染料碘化己锭（HI）和SYTO 13联合用作悬浮液中未固定微生物的革兰氏染色剂。HI可穿透革兰氏阳性菌，但不能穿透革兰氏阴性菌，而SYTO 13可穿透两者。当两种染料一起使用时，SYTO 13使革兰氏阴性菌发出绿色荧光；相反，HI使革兰氏阳性菌发出红橙色荧光，同时淬灭SYTO 13的绿色荧光。该技术正确预测了45株临床相关微生物的革兰氏状态，包括几种已知革兰氏染色不定的菌株。此外，革兰氏阳性厌氧微生物的代表性菌株，在传统革兰氏染色过程中通常会脱色，该方法也能将其正确分类。
