Repetitive-PCR (rep-PCR) using BOXA1R and BOXA2R as single primers was investigated for its potential to genotype bacteriophage. Previously, this technique has been primarily used for the discrimination of bacterial strains. Reproducible DNA fingerprint patterns for various phage types were generated using either of the two primers. The similarity index of replicates ranged from 89.4–100% for BOXA2R-PCR, and from 90 to 100% for BOXA1R-PCR. The method of DNA isolation (p = 0.08) and the phage propagation conditions at two different temperatures (p = 0.527) had no significant influence on generated patterns. Rep-PCR amplification products were generated from different templates including purified phage DNA, phage lysates and phage plaques. The use of this method enabled comparisons of phage genetic profiles to establish their similarity to related or unrelated phages and their bacterial hosts. The findings suggest that repetitive-PCR could be used as a rapid and inexpensive method to preliminary screen phage isolates prior to their selection for more comprehensive studies. The adoption of this rapid, simple and reproducible technique could facilitate preliminary characterisation of a large number of phage isolates and the investigation of genetic relationship between phage genotypes.

中文翻译：

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使用 BOXA 重复 PCR 进行噬菌体基因分型。


使用 BOXA1R 和 BOXA2R 作为单一引物的重复 PCR (rep-PCR) 研究了其对噬菌体进行基因分型的潜力。此前，该技术主要用于细菌菌株的鉴别。使用这两种引物中的任一种都可以生成各种噬菌体类型的可重复 DNA 指纹图谱。 BOXA2R-PCR 的重复相似性指数为 89.4-100%，BOXA1R-PCR 的重复相似性指数为 90-100%。 DNA 分离方法 (p = 0.08) 和两个不同温度下的噬菌体繁殖条件 (p = 0.527) 对生成的模式没有显着影响。 Rep-PCR 扩增产物由不同模板产生，包括纯化的噬菌体 DNA、噬菌体裂解物和噬菌斑。使用这种方法可以比较噬菌体遗传图谱，以确定它们与相关或不相关噬菌体及其细菌宿主的相似性。研究结果表明，在选择噬菌体分离株进行更全面的研究之前，重复 PCR 可以作为一种快速且廉价的方法来初步筛选噬菌体分离株。采用这种快速、简单且可重复的技术可以促进大量噬菌体分离株的初步表征以及噬菌体基因型之间遗传关系的研究。