ABSTRACT
Infective endocarditis (IE) is a life-threatening disease that is associated with high morbidity and mortality. Its long-term prognosis strongly depends on a timely and optimized antibiotic treatment. Therefore, identification of the causative pathogen is crucial and currently based on blood cultures followed by characterization and susceptibility testing of the isolate. However, antibiotic treatment starting prior to blood sampling or IE caused by fastidious or intracellular microorganisms may cause negative culture results. Here we investigate the additional diagnostic value of broad-range PCR in combination with direct sequencing on resected heart tissue or swabs in patients with tissue or swab culture-negative IE in a routine clinical setting. Sensitivity, specificity, and positive and negative predictive values of broad-range PCR from diagnostic material in our patients were 33.3%, 76.9%, 90.9%, and 14.3%, respectively. We identified a total of 20 patients (21.5%) with tissue or culture-negative IE who profited by the additional application of broad-range PCR. We conclude that broad-range PCR on resected heart tissue or swabs is an important complementary diagnostic approach. It should be seen as an indispensable new tool for both the therapeutic and diagnostic management of culture-negative IE and we thus propose its possible inclusion in Duke's diagnostic classification scheme.
Subject(s)
DNA, Ribosomal/genetics , Endocarditis/drug therapy , Endocarditis/genetics , RNA, Ribosomal, 16S/genetics , Aged , Anti-Bacterial Agents/therapeutic use , Bacteria/genetics , Bacteria/isolation & purification , Bacteria/pathogenicity , Blood Culture , DNA, Ribosomal/isolation & purification , Endocarditis/microbiology , Endocarditis/surgery , Female , High-Throughput Nucleotide Sequencing , Humans , Male , Middle Aged , Myocardium/metabolism , Myocardium/pathology , RNA, Ribosomal, 16S/isolation & purification , Thoracic SurgerySubject(s)
Acinetobacter Infections/microbiology , Acinetobacter baumannii/drug effects , Acinetobacter baumannii/enzymology , Bacterial Proteins/genetics , beta-Lactam Resistance , beta-Lactamases/genetics , Acinetobacter baumannii/classification , Acinetobacter baumannii/genetics , Anti-Bacterial Agents/pharmacology , Genotype , Humans , Microbial Sensitivity Tests , Molecular TypingABSTRACT
Latest major contributions in the field of sepsis diagnostics result from advances in PCR technologies permitting new standards in speed and quality, given the fact that a timely diagnosis is the decisive factor to the survival of patients with bloodstream infections.Multiplex real-time PCR is a quantitative method for simultaneous amplification and detection of different targeted DNA molecules within hours. Nevertheless, various studies have shown a number of technical shortcomings as well as a high heterogeneity in sensitivity.The present method allows the standardized and rapid detection and identification of 25 common bacteria and fungi responsible for bloodstream infections from whole blood samples by using LightCycler(®) SeptiFast (LC-SF) test, based on real-time PCR.
