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首页> 外文期刊>Journal of Clinical Microbiology >PCR primers and probes for the 16S rRNA gene of most species of pathogenic bacteria, including bacteria found in cerebrospinal fluid.
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PCR primers and probes for the 16S rRNA gene of most species of pathogenic bacteria, including bacteria found in cerebrospinal fluid.

机译：大多数病原细菌（包括脑脊髓液中发现的细菌）16S rRNA基因的PCR引物和探针。

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A set of broad-range PCR primers for the 16S rRNA gene in bacteria were tested, along with three series of oligonucleotide probes to detect the PCR product. The first series of probes is broad in range and consists of a universal bacterial probe, a gram-positive probe, a Bacteroides-Flavobacterium probe, and two probes for other gram-negative species. The second series was designed to detect PCR products from seven major bacterial species or groups frequently causing meningitis: Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, S. agalactiae, Escherichia coli and other enteric bacteria, Listeria monocytogenes, and Staphylococcus aureus. The third series was designed for the detection of DNA from species or genera commonly considered potential contaminants of clinical samples, including cerebrospinal fluid (CSF): Bacillus, Corynebacterium, Propionibacterium, and coagulase-negative Staphylococcus spp. The primers amplified DNA from all 124 different species of bacteria tested. Southern hybridization testing of the broad-range probes with washes containing 3 M tetramethylammonium chloride indicated that this set of probes correctly identified all but two of the 102 bacterial species tested, the exceptions being Deinococcus radiopugnans and Gardnerella vaginalis. The gram-negative and gram-positive probes hybridized to isolates of two newly characterized bacteria, Alloiococcus otitis and Rochalimaea henselii, as predicted by Gram stain characteristics. The CSF pathogen and contaminant probe sequences were compared with available sequence information and with sequencing data for 32 different species. Testing of the CSF pathogen and contaminant probes against DNA from over 60 different strains indicated that, with the exception of the coagulase-negative Staphylococcus probes, these probes provided the correct identification of bacterial species known to be found in CSF.
机译：测试了一组用于细菌中16S rRNA基因的宽范围PCR引物，以及三个系列的寡核苷酸探针以检测PCR产物。第一组探针的范围很广，由通用细菌探针，革兰氏阳性探针，拟杆菌-黄杆菌探针和两个其他革兰氏阴性菌探针组成。第二个系列旨在检测经常引起脑膜炎的七个主要细菌物种或组的PCR产物：脑膜炎奈瑟菌，流感嗜血杆菌，肺炎链球菌，无乳链球菌，大肠杆菌和其他肠细菌，单核细胞增生李斯特菌和金黄色葡萄球菌。第三系列旨在检测通常被认为是临床样品潜在污染物的物种或属中的DNA，包括脑脊液（CSF）：芽孢杆菌，棒状杆菌，丙酸杆菌和凝固酶阴性葡萄球菌属。引物从测试的所有124种不同细菌中扩增DNA。使用含有3M四甲基氯化铵的洗涤液对宽范围探针进行的Southern杂交测试表明，这套探针正确地鉴定了所测试的102种细菌中的两种细菌，但其中只有两种，例外是放线杆菌和阴道加德纳菌。革兰氏阴性和革兰氏阳性探针可与革兰氏染色特征所预测的两种新近鉴定的细菌（Alloiococcus otitis和Rochalimaea henselii）的细菌杂交。将CSF病原体和污染物探针序列与可用序列信息和32种不同物种的测序数据进行了比较。对CSF病原体和污染物探针针对来自60多个不同菌株的DNA的测试表明，除凝固酶阴性葡萄球菌探针外，这些探针可正确鉴定已知在CSF中发现的细菌种类。

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