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用于土壤法医比对的多重末端限制性片段长度多态性微生物DNA分析及样品条件的影响

Microbial DNA profiling by multiplex terminal restriction fragment length polymorphism for forensic comparison of soil and the influence of sample condition.

作者信息

Macdonald L M, Singh B K, Thomas N, Brewer M J, Campbell C D, Dawson L A

机构信息

The Macaulay Institute, Craigiebuckler, Aberdeen, UK.

出版信息

J Appl Microbiol. 2008 Sep;105(3):813-21. doi: 10.1111/j.1365-2672.2008.03819.x. Epub 2008 Apr 19.

DOI:10.1111/j.1365-2672.2008.03819.x
PMID:18429978
Abstract

AIMS

To evaluate: (i) the impact of air-drying on bacterial, archaeal and fungal soil DNA profiles and (ii) the potential use of multiplex-terminal restriction fragment length polymorphism (M-TRFLP) as a tool for forensic comparison of soil.

METHODS AND RESULTS

An M-TRFLP approach was used to profile bacterial, archaeal and fungal DNA profiles from five different soil sites. Air-drying soil significantly reduced the quantity of DNA but the number of operational taxanomic units (OTU) was unaffected. The impact of air-drying on soil DNA profiles was dependent on soil site and microbial primers. Fungal profiles were altered the least by air-drying. For prokaryotic profiles, air-drying altered the relative similarity/dissimilarity between soil sites. The M-TRFLP approach was more discriminatory compared with soil colour and single-taxa profiling, but did not significantly improve resolution between two similar soils.

CONCLUSIONS

Of those tested, soil fungi were potentially the more robust target for application to soil forensic studies as they were altered less by air-drying and provided clear discrimination of soils from different sites. The M-TRFLP method demonstrated potential to achieve greater resolution, discriminating the soil sites based on both bacterial and fungal components.

SIGNIFICANCE AND IMPACT OF THE STUDY

Soil DNA profiling has potential as a forensic tool, but sample condition and the appropriate selection of microbial target taxa must be considered.

摘要

目的

评估:(i)风干对土壤细菌、古菌和真菌DNA图谱的影响,以及(ii)多重末端限制性片段长度多态性(M-TRFLP)作为土壤法医比对工具的潜在用途。

方法与结果

采用M-TRFLP方法分析来自五个不同土壤地点的细菌、古菌和真菌DNA图谱。风干土壤显著减少了DNA的数量,但操作分类单元(OTU)的数量未受影响。风干对土壤DNA图谱的影响取决于土壤地点和微生物引物。风干对真菌图谱的改变最小。对于原核生物图谱,风干改变了土壤地点之间的相对相似性/不相似性。与土壤颜色和单一分类单元分析相比,M-TRFLP方法具有更高的鉴别力,但在区分两种相似土壤时并没有显著提高分辨率。

结论

在测试的对象中,土壤真菌可能是应用于土壤法医研究的更可靠目标,因为它们受风干的影响较小,并且能够清晰地区分不同地点的土壤。M-TRFLP方法显示出实现更高分辨率的潜力,能够基于细菌和真菌成分区分土壤地点。

研究的意义和影响

土壤DNA分析有潜力作为一种法医工具,但必须考虑样本条件和微生物目标分类单元的适当选择。

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