Kiehlbauch J A, Cameron D N, Wachsmuth I K
Enteric Diseases Branch, Centers for Disease Control and Prevention, Atlanta, GA 30333.
Mol Cell Probes. 1994 Apr;8(2):109-15. doi: 10.1006/mcpr.1994.1015.
Restriction fragment length polymorphisms of ribosomal DNA (ribotyping) of many bacterial species has been useful for both epidemiologic subtyping and species identification. However, the use of ribotyping has been confined to major research and reference laboratories due to two factors: (a) the procedure must be carefully optimized for each organism one wishes to investigate and (b) most currently available protocols use hazardous chemicals or radioisotopes. The purpose of this study is to suggest an overall scheme that a clinical or research microbiologist could apply to ribotyping of any organism. In general, we recommend using a guanidium extraction method for DNA extraction, careful optimization of restriction conditions, and hybridization with non-radioactive digoxigenin-labelled probes; these procedures do not use hazardous chemicals or radioisotopes.
许多细菌物种的核糖体DNA限制性片段长度多态性分析（核糖体分型）在流行病学亚型分析和物种鉴定方面都很有用。然而，由于两个因素，核糖体分型的应用仅限于主要的研究和参考实验室：（a）必须针对每种想要研究的生物体仔细优化该程序；（b）目前大多数可用的方案都使用危险化学品或放射性同位素。本研究的目的是提出一种总体方案，临床或研究微生物学家可将其应用于任何生物体的核糖体分型。一般来说，我们建议使用胍提取法进行DNA提取，仔细优化限制性内切酶反应条件，并与非放射性地高辛标记探针进行杂交；这些程序不使用危险化学品或放射性同位素。
