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首页> 外文期刊>Journal of Clinical Microbiology >Clinical application of novel sample processing technology for the identification of salmonellae by using DNA probes.
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Clinical application of novel sample processing technology for the identification of salmonellae by using DNA probes.

机译：利用DNA探针鉴定沙门氏菌的新型样品处理技术的临床应用。

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Two hundred and fifty clinical fecal specimens collected over a 7-month period were analyzed for the presence of salmonellae by a rapid DNA hybridization procedure. Hybridizations were performed by using a novel specimen processing protocol called wicking and a previously unreported 1,600-base-pair probe cloned from Salmonella enteritidis DNA. The probe was shown to be reactive with all 70 Salmonella serotypes tested and not reactive with 101 stock strains of other enteric bacteria. Southern analysis of 30 Salmonella isolates representing 22 serotypes suggested that the probe sequence was highly conserved, appearing as a 1,600-base-pair band in a BglII digest of isolate DNA in 29 of 30 isolates and as a 2,300-base-pair fragment in 1 of the isolates. The probe correctly identified all salmonellae (nine isolates) among 47 H2S-producing colonies tested from among 250 clinical specimens cultured on xylose-lysine-desoxycholate medium. Salmonellae grown on xylose-lysine-desoxycholate medium gave consistently higher hybridization values than did those grown on either MacConkey or Hektoen enteric agar. In addition, of eight gram-negative broth enrichments in which salmonellae were identified by conventional means, seven were probe positive. The use of this nucleic acid probe and hybridization technique provides a simple and rapid identification of Salmonella species.
机译：在7个月的时间内收集的250个临床粪便标本通过快速DNA杂交程序分析了沙门氏菌的存在。杂交是通过使用一种称为“芯吸”的新型标本处理方案以及从肠炎沙门氏菌DNA克隆的先前未报道的1,600个碱基对的探针进行的。该探针与所有70种沙门氏菌血清型均具有反应性，与101种其他肠细菌的原种无反应。对代表22种血清型的30种沙门氏菌分离株进行的Southern分析表明，该探针序列高度保守，在30种分离株中的29种分离株DNA的BglII消化物中显示为1,600个碱基对的条带，在1种中显示为2,300个碱基对的片段分离株。该探针正确鉴定了木糖-赖氨酸-脱氧胆酸盐培养基上培养的250个临床标本中47个产生H2S的菌落中的所有沙门氏菌（九个分离株）。在木糖-赖氨酸-脱氧胆酸盐培养基上生长的沙门氏菌比在MacConkey或Hektoen肠琼脂上生长的沙门氏菌始终具有更高的杂交值。此外，在通过常规方法鉴定出沙门氏菌的八种革兰氏阴性肉汤浓缩物中，有七种探针阳性。该核酸探针和杂交技术的使用提供了沙门氏菌种类的简单和快速鉴定。

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