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Applied and Environmental Microbiology, November 2000, p. 5066-5072, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Rapid Detection and Quantification of Members of the Archaeal Community by Quantitative PCR Using Fluorogenic Probes

Ken Takai* and Koki Horikoshi

Deep-sea Microorganisms Research Group, Japan Marine Science & Technology Center, Yokosuka 237-0061, Japan

Received 1 June 2000/Accepted 14 August 2000

We describe a rapid, reproducible, and sensitive method for detection and quantification of archaea in naturally occurring microbial communities. A domain-specific PCR primer set and a domain-specific fluorogenic probe having strong and weak selectivity, respectively, for archaeal rRNA genes (rDNAs) were designed. A universal PCR primer set and a universal fluorogenic probe for both bacterial and archaeal rDNAs were also designed. Using these primers and probes, we demonstrated that detection and quantification of archaeal rDNAs in controlled microbial rDNA assemblages can be successfully achieved. The system which we designed was also able to detect and quantify archaeal rDNAs in DNA samples obtained not only from environments in which thermophilic archaea are abundant but also from environments in which methanogenic archaea are abundant. Our findings indicate that this method is applicable to culture-independent molecular analysis of microbial communities in various environments.

* Corresponding author. Mailing address: Deep-sea Microorganisms Research Group, Japan Marine Science & Technology Center, 2-15 Natsushima-cho, Yokosuka 237-0061, Japan. Phone: 81-468-67-3894. Fax: 81-468-66-6364. E-mail: kent{at}jamstec.go.jp.

Applied and Environmental Microbiology, November 2000, p. 5066-5072, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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