通过rRNA与DNA乳胶杂交检测细菌以及对杂交体进行免疫检测。
Detection of bacteria by hybridization of rRNA with DNA-latex and immunodetection of hybrids.
作者信息
Miller C A, Patterson W L, Johnson P K, Swartzell C T, Wogoman F, Albarella J P, Carrico R J
机构信息
Miles, Inc., Elkhart, Indiana 46515.
出版信息
J Clin Microbiol. 1988 Jul;26(7):1271-6. doi: 10.1128/jcm.26.7.1271-1276.1988.
A novel nucleic acid hybridization assay with a DNA probe immobilized on 1.25-micron-diameter latex particles was developed. Hybridization of the immobilized probe DNA with sample rRNA was complete in 10 to 15 min. Alkaline phosphatase-labeled anti-DNA-RNA was allowed to bind to the DNA-RNA hybrids on the latex particles. Then the latex was collected on a small glass fiber filter pad, and bound alkaline phosphatase was quantitated by reflectance rate measurement. The method detected a broad range of bacterial species and had a detection limit of 500 cells per assay. The assay was used to screen urine samples for bacteriuria and had a sensitivity of 96.2% compared with conventional culture at a decision level of greater than or equal to 10(4) CFU/ml. The hybridization method could have broad application to the detection of bacteria and viruses.
开发了一种新型核酸杂交测定法，其使用固定在直径1.25微米乳胶颗粒上的DNA探针。固定化探针DNA与样品rRNA的杂交在10至15分钟内完成。使碱性磷酸酶标记的抗DNA-RNA与乳胶颗粒上的DNA-RNA杂交体结合。然后将乳胶收集在小玻璃纤维滤垫上，并通过反射率测量对结合的碱性磷酸酶进行定量。该方法可检测多种细菌种类，每次测定的检测限为500个细胞。该测定法用于筛查尿液样本中的菌尿，在判定水平大于或等于10(4) CFU/ml时，与传统培养法相比灵敏度为96.2%。该杂交方法可广泛应用于细菌和病毒的检测。
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