Advantages and Limitations of Direct PCR Amplification of Bacterial 16S-rDNA from Resected Heart Tissue or Swabs Followed by Direct Sequencing for Diagnosing Infective Endocarditis: A Retrospective Analysis in the Routine Clinical Setting.
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.
Institute of Laboratory Medicine, Microbiology and Infection Control, The Northwest Medical Centre, 60488 Frankfurt am Main, Germany.
Daniela
D
Maneg
Institute of Laboratory Medicine, Microbiology and Infection Control, The Northwest Medical Centre, 60488 Frankfurt am Main, Germany.
Janina
J
Sponsel
Institute of Laboratory Medicine, Microbiology and Infection Control, The Northwest Medical Centre, 60488 Frankfurt am Main, Germany.
Iris
I
Müller
Institute of Laboratory Medicine, Microbiology and Infection Control, The Northwest Medical Centre, 60488 Frankfurt am Main, Germany.
Benedikt
B
Lohr
CAPHRI School for Public Health and Primary Care, Department of Medical Microbiology, Maastricht University Medical Center, 6200 MD Maastricht, Netherlands.
John
J
Penders
Kerckhoff Klinik, 61231 Bad Nauheim, Germany.
Katharina
K
Madlener
Institute of Laboratory Medicine, Microbiology and Infection Control, The Northwest Medical Centre, 60488 Frankfurt am Main, Germany.
Klaus-Peter
KP
Hunfeld
D000368:Aged; D000900:Anti-Bacterial Agents / Q000627:therapeutic use; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D000071997:Blood Culture; D004275:DNA, Ribosomal / Q000235:genetics* / Q000302:isolation & purification; D004696:Endocarditis / Q000188:drug therapy* / Q000235:genetics* / Q000382:microbiology / Q000601:surgery; D005260:Female; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008875:Middle Aged; D009206:Myocardium / Q000378:metabolism / Q000473:pathology; D012336:RNA, Ribosomal, 16S / Q000235:genetics* / Q000302:isolation & purification; D013903:Thoracic Surgery
D000900:Anti-Bacterial Agents; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S