用于分子检测、确认和鉴定梭杆菌属细菌的属特异性聚合酶链反应检测方法的开发

Development of a genus-specific PCR assay for the molecular detection, confirmation and identification of Fusobacterium spp.

作者信息

Nagano Y, Watabe M, Porter K G, Coulter W A, Millar B C, Elborn J S, Goldsmith C E, Rooney P J, Loughrey A, Moore J E

机构信息

Northern Ireland Public Health Laboratory, Department of Bacteriology, Belfast City Hospital, UK.

出版信息

Br J Biomed Sci. 2007;64(2):74-7. doi: 10.1080/09674845.2007.11732760.

DOI:10.1080/09674845.2007.11732760

PMID:17633142

Abstract

A genus-specific polymerase chain reaction (PCR)-based assay is developed for the detection and identification of clinically relevant Fusobacterium species, including F. nucleatum and F. necrophorum. Two 16S ribosomal DNA (rDNA) primers, FUSO1 (forward primer: 5'-GAG AGA GCT TTG CGT CC-3' [17-mer]) and FUSO 2 (reverse primer: 5'-TGG GCG CTG AGG TTC GAC -3' [18-mer]) are designed to target conserved regions of the 16S rDNA gene for Fusobacterium spp. Subsequent proof-of-principle studies employing this assay detected Fusobacterium spp. in the faeces of eight (10%) out of 80 patients with suspected gastrointestinal infection. This assay may be used for the genus-specific detection of Fusobacterium spp. from clinical specimens and for subsequent species identification.

摘要

开发了一种基于属特异性聚合酶链反应（PCR）的检测方法，用于检测和鉴定临床相关的梭杆菌属菌种，包括具核梭杆菌和坏死梭杆菌。设计了两条16S核糖体DNA（rDNA）引物，FUSO1（正向引物：5'-GAG AGA GCT TTG CGT CC-3'[17个碱基对]）和FUSO 2（反向引物：5'-TGG GCG CTG AGG TTC GAC -3'[18个碱基对]），以靶向梭杆菌属16S rDNA基因的保守区域。随后采用该检测方法的原理验证研究在80例疑似胃肠道感染患者中的8例（10%）粪便中检测到了梭杆菌属菌种。该检测方法可用于从临床标本中对梭杆菌属菌种进行属特异性检测，并用于后续的菌种鉴定。