Shangkuan Y H, Lin H C
Division of Bacteriology, Institute of Preventive Medicine, National Defense Medical Center, Sanhsia Taipei, Taiwan, Republic of China.
J Appl Microbiol. 1998 Oct;85(4):693-702. doi: 10.1111/j.1365-2672.1998.00582.x.
A Random Amplified Polymorphic DNA (RAPD) fingerprinting method was developed to differentiate isolates of Salmonella serotype typhi (S. typhi) and other Salmonella isolates. A panel of five primers was used to examine 63 isolates of Salm. typhi, including 56 strains isolated in Taiwan and seven strains obtained abroad. Twenty-one RAPD types were revealed using the RAPD fingerprinting method. An RAPD with primer 6032 yielded a polymorphism in a 350 bp fragment that differentiated the attenuated vaccine strain Salm. typhi Ty21a from the rest of the Salm. typhi strains. Strains of Salm. typhi were divided into five types with primer D14307. Primer D14307 also proved capable of discrimination among 65 other Salmonella isolates representing 42 different serotypes. The bacterial DNA used in this RAPD protocol was obtained using a commercially available DNA extraction kit (GeneReleaser). The DNA of various strains of Salmonella from this simple extraction procedure could be discriminated within a few hours using the RAPD technique.
开发了一种随机扩增多态性DNA（RAPD）指纹图谱方法，以区分伤寒沙门氏菌（S. typhi）分离株和其他沙门氏菌分离株。使用一组五种引物来检测63株伤寒沙门氏菌分离株，其中包括在台湾分离的56株菌株和从国外获得的7株菌株。使用RAPD指纹图谱方法揭示了21种RAPD类型。用引物6032进行的RAPD在一个350 bp片段中产生了多态性，该片段将减毒疫苗株伤寒沙门氏菌Ty21a与其余伤寒沙门氏菌菌株区分开来。用引物D14307将伤寒沙门氏菌菌株分为五种类型。引物D14307也被证明能够区分代表42种不同血清型的65株其他沙门氏菌分离株。本RAPD方案中使用的细菌DNA是使用市售DNA提取试剂盒（GeneReleaser）获得的。使用RAPD技术，通过这种简单的提取程序获得的各种沙门氏菌菌株的DNA可以在几小时内得到区分。
