Bacterial species identification after DNA amplification with a universal primer pair

被引:82
作者
McCabe, KM
Zhang, YH
Huang, BL
Wagar, EA
McCabe, ERB
机构
[1] Univ Calif Los Angeles, Sch Med, Dept Pediat, Los Angeles, CA 90095 USA
[2] Univ Calif Los Angeles, Sch Med, Dept Pathol, Los Angeles, CA 90095 USA
[3] Univ Calif Los Angeles, Sch Med, Dept Lab Med, Los Angeles, CA 90095 USA
[4] Univ Calif Los Angeles, Sch Med, Inst Mol Biol, Los Angeles, CA 90095 USA
关键词
ribosomal RNA; 6S; polymerase chain reaction (PCR); bacterial identification; infectious agents; bacteria; molecular diagnostics; DNA amplification; amplification; DNA;
D O I
10.1006/mgme.1998.2795
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The diagnosis of bacterial infections can be difficult and time consuming. Rapid and reliable molecular triage of potentially infected patients, particularly the young and the elderly, would prevent unnecessary hospitalizations, reduce associated medical costs, and improve the quality of care. Polymerase chain reaction (PCR) amplification utilizing a universal bacterial primer pair, followed by hybridization with species-specific probes, would allow rapid identification of the presence or absence of bacterial DNA, along with an identification of the bacterial species present. Molecular microbiological analyses will require access to bacterial strain standards that can be catalogued and distributed to clinical laboratories. We amplified template DNA in filter paper spots containing boiled bacteria from 14 clinical isolates using a universal primer pair for the 16S ribosomal RNA (rDNA) coding sequence. Species-specific probes were hybridized to the amplification products for bacterial species identification. We conclude that template DNA can be identified with species-specific probes after universal bacterial amplification with a single primer pair. We also demonstrate a rapid and efficient method for the long-term storage and cataloguing of bacterial DNA for use in quality control at clinical laboratories adopting molecular diagnostic methodologies. We speculate that PCR amplification combined with species-specific probe hybridization not only will represent an improvement over culture-based methods in terms of speed, sensitivity, and cost, but will also allow for the identification of unculturable bacteria and emerging or reemerging pathogenic organisms. (C) 1999 Academic Press.
引用
收藏
页码:205 / 211
页数:7
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