UNSPECIFIED. (2003) Development and validation of a diagnostic microbial microarray for methanotrophs. ENVIRONMENTAL MICROBIOLOGY, 5 (7). pp. 566-582. ISSN 1462-2912

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Abstract

The potential of DNA microarray technology in high-throughput detection of bacteria and quantitative assessment of their community structures is widely acknowledged but has not been fully realised yet. A generally applicable set of techniques, based on readily available technologies and materials, was developed for the design, production and application of diagnostic microbial microarrays. A microarray targeting the particulate methane monooxygenase (pmoA ) gene was developed for the detection and quantification of methanotrophs and functionally related bacteria. A microarray consisting of a set of 59 probes that covers the whole known diversity of these bacteria was validated with a representative set of extant strains and environmental clones. The potential of the pmoA microarray was tested with environmental samples. The results were in good agreement with those of clone library sequence analyses. The approach can currently detect less dominant bacteria down to 5% of the total community targeted. Initial tests assessing the quantification potential of this system with artificial PCR mixtures showed very good correlation with the expected results with standard deviations in the range of 0.4-17.2%. Quantification of environmental samples with this method requires the design of a reference mixture consisting of very close relatives of the strains within the sample and is currently limited by biases inherent in environmental DNA extraction and universal PCR amplification.

Item Type:	Journal Article
Subjects:	Q Science > QR Microbiology
Journal or Publication Title:	ENVIRONMENTAL MICROBIOLOGY
Publisher:	BLACKWELL PUBLISHING LTD
ISSN:	1462-2912
Official Date:	July 2003
Volume:	5
Number:	7
Number of Pages:	17
Page Range:	pp. 566-582
Identification Number:	10.1046/j.1462-2920.2003.00450.x
Publication Status:	Published
URI:	http://wrap.warwick.ac.uk/id/eprint/9618

Data sourced from Thomson Reuters' Web of Knowledge

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