Group‐specific primer and probe sets to detect methanogenic communities using quantitative real‐time polymerase chain reaction

Biotechnology and Bioengineering - Tập 89 Số 6 - Trang 670-679 - 2005
Youngseob Yu1,2,3, Changsoo Lee1,2,3, Jaai Kim1,2,3, Seokhwan Hwang1,2,3
1School of Environmental Science and Engineering, Pohang University of Science and Technology, Pohang, Kyungbuk 790-784, South Korea
2fax: +8254-279-8299
3telephone: +8254-279-2282

Tóm tắt

AbstractReal‐time polymerase chain reaction (PCR) is a highly sensitive method that can be used for the detection and quantification of microbial populations without cultivating them in anaerobic processes and environmental samples. This work was conducted to design primer and probe sets for the detection of methanogens using a real‐time PCR with the TaqMan system. Six group‐specific methanogenic primer and probe sets were designed. These sets separately detect four orders (Methanococcales, Methanobacteriales, Methanomicrobiales, and Methanosarcinales) along with two families (Methanosarcinaceae and Methanosaetaceae) of the order Methanosarcinales. We also designed the universal primer and probe sets that specifically detect the 16S rDNA of prokaryotes and of the domain Bacteria and Archaea, and which are fully compatible with the TaqMan real‐time PCR system. Target‐group specificity of each primer and probe set was empirically verified by testing DNA isolated from 28 archaeal cultures and by analyzing potential false results. In general, each primer and probe set was very specific to the target group. The primer and probe sets designed in this study can be used to detect and quantify the order‐level (family‐level in the case of Methanosarcinales) methanogenic groups in anaerobic biological processes and various environments. © 2005 Wiley Periodicals, Inc.

Từ khóa


Tài liệu tham khảo

10.1111/j.1574-6941.2006.00225.x

10.1128/mr.59.1.143-169.1995

10.1093/nar/gkf450

Atlas RM, 1993, Microbial ecology: fundamentals and applications

10.1007/978-1-4615-2391-8_2

ChunJ.1995.Computer‐assisted classification and identification of actinomycetes. PhD thesis University of Newcastle upon Tyne. Newcastle upon Tyne UK.

10.1093/nar/gkg039

10.1128/JCM.40.6.2224-2227.2002

Garrity GM, 2001, Phylum AI. Crenarchaeota phy. nov, 169

Garrity GM, 2001, Phylum AII. Euryarchaeota phy. nov, 211

Grady CPL, 1999, Biological wastewater treatment, 1076

10.1002/(SICI)1097-0290(19980205)57:3<342::AID-BIT11>3.0.CO;2-I

10.1128/JCM.40.7.2392-2397.2002

10.1128/AEM.67.7.3122-3126.2001

10.1093/clinchem/47.6.1124

10.1016/S0958-1669(98)80082-7

10.1016/S0043-1354(01)00377-3

10.1101/gr.4.6.357

Madigan MT, 2000, Brock: biology of microorganisms, 545

10.1128/JCM.40.5.1698-1704.2002

10.1016/S0043-1354(98)00454-0

10.1099/00221287-148-1-257

10.1023/A:1008300423935

10.1007/978-1-4757-2191-1_44

Raskin L, 1994, Quantificaiton of methanogenic groups in anaerobic biological reactors by oligonuclotide probe hybridization, Appl Environ Microbiol, 60, 1241, 10.1128/aem.60.4.1241-1248.1994

10.1128/AEM.60.4.1232-1240.1994

10.1007/BF00874140

Reischl U, 2002, Rapid cycle real‐time PCR—methods and applications: microbiology and food anaysis, 10.1007/978-3-642-48351-6

10.1128/AEM.58.10.3417-3418.1992

Rittmann BE, 2001, Environmental biotechnology: principles and applications

10.1016/S0958-1669(00)00210-X

Sowers KR, 1995, Archaea: a laboratory manual—methanogens, 459

Speece RE, 1996, Anaerobic biotechnology for industrial wastewaters, 394

10.1128/AEM.66.11.4605-4614.2000

10.1128/AEM.66.11.5066-5072.2000

Vogels GD, 1988, Biology of anaerobic microorganisms, 707

10.1128/AEM.67.9.3985-3993.2001

10.1006/meth.2001.1265

10.1073/pnas.87.12.4576

10.1016/S0043-1354(03)00006-X

10.1002/bit.10164

10.1007/978-1-4615-2391-8_4

Thông tin
Thông tin xuất bản

Biotechnology and Bioengineering

Tập 89 Số 6

670-679

Thông tin tác giả