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首页> 外文期刊>Applied Microbiology >Genotyping of Present-Day and Historical Geobacillus Species Isolates from Milk Powders by High-Resolution Melt Analysis of Multiple Variable-Number Tandem-Repeat Loci
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Genotyping of Present-Day and Historical Geobacillus Species Isolates from Milk Powders by High-Resolution Melt Analysis of Multiple Variable-Number Tandem-Repeat Loci

机译：多个可变数串联重复基因座的高分辨率熔解分析法对奶粉中现今和历史地理杆菌种的基因分型

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Spores of thermophilic Geobacillus species are a common contaminant of milk powder worldwide due to their ability to form biofilms within processing plants. Genotyping methods can provide information regarding the source and monitoring of contamination. A new genotyping method was developed based on multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) in conjunction with high-resolution melt analysis (MLV-HRMA) and compared to the currently used method, randomized amplified polymorphic DNA PCR (RAPD-PCR). Four VNTR loci were identified and used to genotype 46 Geobacillus isolates obtained from retailed powder and samples from 2 different milk powder processing plants. These 46 isolates were differentiated into 16 different groups using MLV-HRMA ( D = 0.89). In contrast, only 13 RAPD-PCR genotypes were identified among the 46 isolates ( D = 0.79). This new method was then used to analyze 35 isolates obtained from powders with high spore counts (>10~(4) spores · g~(?1)) from a single processing plant together with 27 historical isolates obtained from powder samples processed in the same region of Australia 17 years ago. Results showed that three genotypes can coexist in a single processing run, while the same genotypes observed 17 years ago are present today. While certain genotypes could be responsible for powders with high spore counts, there was no correlation to specific genotypes being present in powder plants and retailed samples. In conclusion, the MLV-HRMA method is useful for genotyping Geobacillus spp. to provide insight into the prevalence and persistence of certain genotypes within milk powder processing plants.
机译：嗜热地芽孢杆菌属的孢子由于在加工厂内形成生物膜的能力而成为全世界奶粉的常见污染物。基因分型方法可以提供有关污染来源和监测的信息。基于多基因座可变数目串联重复（VNTR）分析（MLVA）结合高分辨率熔解分析（MLV-HRMA）研发了一种新的基因分型方法，并将其与当前使用的方法进行了比较，即随机扩增多态性DNA PCR（RAPD） -PCR）。鉴定出四个VNTR基因座，并将其用于对从零售奶粉和来自2个不同奶粉加工厂的样品中获得的46种Geobacillus菌株进行基因分型。使用MLV-HRMA将这46个分离株分为16个不同的组（D = 0.89）。相反，在46个分离株中仅鉴定出13个RAPD-PCR基因型（D = 0.79）。然后使用此新方法分析了来自单个加工工厂的高孢子计数（> 10〜（4）孢子·g〜（？1））粉末中的35种分离物以及27种历史分离物，这些历史分离物是从单处理工厂中加工的粉末样品中获得的。 17年前是澳大利亚的同一地区。结果表明，三种基因型可以在一个加工过程中共存，而如今却存在17年前观察到的相同基因型。虽然某些基因型可能是造成高孢子数的粉末的原因，但与粉末植物和零售样本中存在的特定基因型没有关联。总之，MLV-HRMA方法可用于对Geobacillus spp进行基因分型。提供有关奶粉加工厂内某些基因型的普遍性和持久性的见解。

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