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Specific detection of Campylobacter jejuni and Campylobacter coli by using polymerase chain reaction.

机译:通过聚合酶链反应特异性检测空肠弯曲菌和大肠弯曲菌。

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摘要

Development of a routine detection assay for Campylobacter jejuni and Campylobacter coli in clinical specimens was undertaken by using the polymerase chain reaction (PCR). An oligonucleotide primer pair from a conserved 5' region of the flaA gene of C. coli VC167 was used to amplify a 450-bp region by PCR. The primer pair specifically detected 4 strains of C. coli and 47 strains of C. jejuni; but it did not detect strains of Campylobacter fetus, Campylobacter lari, Campylobacter upsaliensis, Campylobacter cryaerophila, Campylobacter butzleri, Campylobacter hyointestinalis, Wolinella recta, Helicobacter pylori, Escherichia coli, Shigella spp., Salmonella spp., Vibrio cholerae, Citrobacter freundii, or Aeromonas spp. By using a nonradioactively labeled probe internal to the PCR product, the assay could detect as little as 0.0062 pg of purified C. coli DNA, or the equivalent of four bacteria. In stools seeded with C. coli cells, the probe could detect between 30 and 60 bacteria per PCR assay. The assay was also successfully used to detect C. coli in rectal swab specimens from experimentally infected rabbits and C. jejuni in human stool samples.
机译:使用聚合酶链反应(PCR)开发了临床标本中空肠弯曲菌和大肠杆菌弯曲杆菌的常规检测方法。来自大肠杆菌VC167的flaA基因的保守5'区域的寡核苷酸引物对用于通过PCR扩增450bp区域。引物对特异性地检测到4株大肠杆状杆菌和47株空肠弯曲杆菌。但它未检测到胎儿弯曲杆菌,拉里弯曲杆菌,upsaliensis弯曲杆菌,低温嗜热弯曲杆菌,Butzleri弯曲杆菌,hypointestinalis弯曲杆菌,Wolinella recta,幽门螺杆菌,大肠杆菌,志贺氏菌,志贺氏菌,沙门氏菌或沙门氏菌。 spp。通过使用PCR产物内部的非放射性标记探针,该检测方法可检测到0.0062 pg纯化的大肠杆菌DNA,或等同于四种细菌。在装有大肠杆菌细胞的粪便中,该探针每次PCR检测可检测30至60个细菌。该测定法还成功用于检测实验感染的兔子的直肠拭子样本中的大肠杆菌和人粪便样本中的空肠弯曲菌。

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