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用于鉴定洋葱伯克霍尔德菌及其与唐菖蒲伯克霍尔德菌鉴别的聚合酶链反应中三种寡核苷酸引物组的评估

Evaluation of three oligonucleotide primer sets in PCR for the identification of Burkholderia cepacia and their differentiation from Burkholderia gladioli.

作者信息

Clode F E, Kaufmann M E, Malnick H, Pitt T L

机构信息

Laboratory of Hospital Infection, Central Public Health Laboratory, London, UK.

出版信息

J Clin Pathol. 1999 Mar;52(3):173-6. doi: 10.1136/jcp.52.3.173.

Abstract

AIMS

To evaluate three oligonucleotide primer pairs--two specific for 16S and 23S rRNA sequences of Burkholderia cepacia, and the third specific for internal transcribed spacer region of 16S-23S sequences of B gladioli--for the identification and differentiation of reference and clinical strains of these and other species.

METHODS

The three primers sets were applied in polymerase chain reaction (PCR) to a collection of 177 clinical isolates submitted for identification from diagnostic laboratories as presumed B cepacia.

RESULTS

At an annealing temperature of 63 degrees C, all eight B cepacia and four B gladioli reference strains reacted with their specific primers. B vandii was the only other species that was positive with both B cepacia primers but five Burkholderia or Ralstonia species reacted with one of these primers. Seventy eight isolates were typical of B cepacia in biochemical tests and 75 of these reacted with specific primers; three, however, were positive with the B gladioli primers. Fifteen asaccharolytic isolates were confirmed as B cepacia by PCR but other non-fermenting Gram negative species were negative with each of the primers.

CONCLUSIONS

PCR using 16S rRNA sequences is recommended for identification of B cepacia that give atypical results in biochemical tests.

摘要

目的

评估三种寡核苷酸引物对——两种针对洋葱伯克霍尔德菌16S和23S rRNA序列的特异性引物对,以及第三种针对唐菖蒲伯克霍尔德菌16S - 23S序列内部转录间隔区的特异性引物对——用于鉴定和区分这些菌种及其他菌种的参考菌株和临床菌株。

方法

将这三种引物组应用于聚合酶链反应(PCR),对从诊断实验室提交鉴定的177株临床分离株进行检测,这些分离株被推测为洋葱伯克霍尔德菌。

结果

在退火温度为63℃时,所有8株洋葱伯克霍尔德菌和4株唐菖蒲伯克霍尔德菌参考菌株均与其特异性引物发生反应。鼻疽伯克霍尔德菌是唯一一种对两种洋葱伯克霍尔德菌引物均呈阳性反应的其他菌种,但有5种伯克霍尔德菌或罗尔斯顿菌属菌种与其中一种引物发生反应。78株分离株在生化试验中表现为典型的洋葱伯克霍尔德菌,其中75株与特异性引物发生反应;然而,有3株对唐菖蒲伯克霍尔德菌引物呈阳性。15株不分解糖的分离株通过PCR被确认为洋葱伯克霍尔德菌,但其他非发酵革兰氏阴性菌种对每种引物均呈阴性。

结论

对于在生化试验中给出非典型结果的洋葱伯克霍尔德菌,建议使用16S rRNA序列的PCR进行鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/348f/501074/a75e3a2aa816/jclinpath00276-0014-a.jpg

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