Complementarity between targeted real-time specific PCR and conventional broad-range 16S rDNA PCR in the syndrome-driven diagnosis of infectious diseases
Complementarity between targeted real-time specific PCR and conventional broad-range 16S rDNA PCR in the syndrome-driven diagnosis of infectious diseases
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DOI：
10.1007/s10096-014-2263-z
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发表时间：
2015-03-01
影响因子：
4.5
通讯作者：
Raoult, D.
中科院分区：
文献类型：
--
作者：
Morel, A-S;Dubourg, G.;Raoult, D.
分子工具在传染病的诊断中显示出附加价值，特别是对那些由挑剔的细胞内微生物引起的疾病，或在取样前接受抗生素治疗的患者。如果16S rDNA扩增在微生物实验室中逐步实施，特异性实时聚合酶链反应（PCR）将允许提高分子方法的灵敏度并减少污染。在此，我们报告了我们两年来在传染病诊断方面的经验，在此期间，我们从法国和国外收到了18,056名患者的32,948份临床样本。共检测81476份，其中阳性1192份。分子技术在31.3%的呼吸道样本、27.8%的心内膜炎样本和51.9%的腺炎样本中检测到细胞内微生物。除胞内细菌外，该系列中25%的阳性样品在培养中无菌。传统的广谱PCR允许鉴定挑剔和厌氧微生物，但特异性实时PCR在诊断骨关节感染，特别是由王氏金氏菌和金黄色葡萄球菌引起的感染，以及心内膜炎诊断，特别是当涉及溶胆链球菌和金黄色葡萄球菌时，显示出显著的优势。传统的广谱PCR对两种技术的总体诊断的敏感性为62.9%。这些发现应该引导微生物学家关注针对临床综合征的靶向特异性实时PCR。最后，综合征驱动诊断包括对每种综合征通常涉及的一组微生物进行检测，允许建立31个偶然诊断。
Molecular tools have shown an added value in the diagnosis of infectious diseases, in particular for those caused by fastidious intracellular microorganisms, or in patients receiving antibiotics before sampling. If 16S rDNA amplification had been gradually implemented in microbiology laboratories, specific real-time polymerase chain reaction (PCR) would have permitted an increase in the sensitivity of molecular methods and a reduction of contamination. Herein, we report our experience in the diagnosis of infectious diseases over two years, during which 32,948 clinical samples from 18,056 patients were received from France and abroad. Among these samples, 81,476 PCRs were performed, of which 1,192 were positive. Molecular techniques detected intracellular microorganisms in 31.3 % of respiratory samples, 27.8 % of endocarditis samples and 51.9 % of adenitis samples. Excluding intracellular bacteria, 25 % of the positive samples in this series were sterile in culture. Conventional broad-range PCR permitted the identification of fastidious and anaerobic microorganisms, but specific real-time PCR showed a significant superiority in the diagnosis of osteoarticular infections, in particular for those caused by Kingella kingae and Staphylococcus aureus, and for endocarditis diagnosis, specifically when Streptococcus gallolyticus and Staphylococcus aureus were involved. The sensitivity of conventional broad-range PCR was 62.9 % concerning overall diagnoses for which both techniques had been performed. These findings should lead microbiologists to focus on targeted specific real-time PCR regarding the clinical syndrome. Finally, syndrome-driven diagnosis, which consists of testing a panel of microorganisms commonly involved for each syndrome, permitted the establishment of 31 incidental diagnoses.