A Modified PCR-Based Method for Rapid Non-Radioactive Detection of Clinically Important Pathogens

• GORELOV Victor N.

  Department of Surgery A, Heinrich Heine University

• DUMON Kristoffel

  Department of Surgery A, Heinrich Heine University

• BARTENEVA Natalia S.

  Department of Surgery A, Heinrich Heine University

• ROHER Hans-Dieter

  Department of Surgery A, Heinrich Heine University

• GORETZKI Peter E.

  Department of Surgery A, Heinrich Heine University

抄録

We have devised a sensitive and rapid method for the detection of several bacterial pathogens in clinical specimens using PCR. This method has been named Direct Labeling and Detection Procedure (DLDP) and is based on the direct incorporation of a nonradioactive digoxigenin label (DIG-11-dUTP) into a microbial species-specific gene fragment during amplification. Following amplification, the resulting PCR products are cleansed of nonincorporated DIG-11-dUTP, spotted onto a nylon membrane, fixed by UV-crosslinking and the labeled DNA is visualized by digoxigenin detection reagents. Using cultivated reference bacteria (Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa) we were able to demonstrate a rapid and sensitive detection of <20 CFU of bacteria in human secretions (sputum, urine, mucous). The present study suggests that DLDP can be used as a reliable method for indication of bacteria in clinical or environmental specimens with the proviso that the selected corresponding oligonucleotide primers provide amplification of strong species-specific genes.

収録刊行物

• Microbiology and immunology

  Microbiology and immunology 40 (9), 611-616, 1996-09-20

  財団法人 学会誌刊行センター

キーワード

• Polymerase chain reaction
• Direct nonradioactive labeling
• Bacterial pathogens

詳細情報

• CRID: 1571417124231855488
• NII論文ID: 10004893843
• NII書誌ID: AA00738350
• ISSN: 03855600
• Web Site: http://chemport.cas.org/cgi-bin/sdcgi?APP=ftslink&action=reflink&origin=jstage2&version=1.0&coi=1:CAS:528:DyaK28XmtlWhtLw%3D&md5=ec977a126021e5eb394d76b420894529
• Web Site: https://search.jamas.or.jp/link/ui/1997042467
• 本文言語コード: en
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