Ann Lab Med.  2017 Nov;37(6):505-510. 10.3343/alm.2017.37.6.505.

Utility of a Direct 16S rDNA PCR and Sequencing for Etiological Diagnosis of Infective Endocarditis

Affiliations
  • 1Department of Laboratory Medicine, Asan Medical Center and University of Ulsan College of Medicine, Seoul, Korea. sung@amc.seoul.kr
  • 2Department of Internal Medicine, Asan Medical Center and University of Ulsan College of Medicine, Seoul, Korea.
  • 3Department of Thoracic and Cardiovascular Surgery, Asan Medical Center and University of Ulsan College of Medicine, Seoul, Korea.

Abstract

BACKGROUND
Cases of infective endocarditis (IE) require prompt etiological diagnosis for effective treatment. Molecular methods can aid in rapid and reliable diagnosis of culture-negative IE cases. We evaluated the utility of 16S rDNA PCR and sequencing in determining the causative agents of IE in valve tissues, especially when specimens were obtained after initiation of antimicrobial therapy.
METHODS
We performed 16S rDNA PCR and sequencing in heart valve specimens and medical records review of 80 patients who underwent protocol-based cardiac surgery from 2013 to 2015. One patient did not meet the criteria for IE. Sixty-five (81.3%) and 14 pa-tients (17.5%) were diagnosed as having definite IE and possible IE, respectively. Blood and heart valve biopsy tissue were examined by using routine microbiological methods.
RESULTS
Blood cultures in our hospital were IE-positive for 33 patients (41.8%), whereas 49 patients (62.0%) showed positive blood cultures when initial blood cultures performed at the referring hospital were included. Eighteen (22.8%) and 40 patients (50.6%) were IE-positive in valve tissue cultures and 16S rDNA PCR, respectively. Bacteria in the Streptococcus mitis group (n=26) were the most common etiological agents of IE. Eight (10.1%) culture-negative specimens tested positive by 16S rDNA PCR. In five of eight PCR-positive and culture-negative cases, fastidious or anaerobic organisms were the cause of IE.
CONCLUSIONS
Direct 16S rDNA PCR and sequencing can be used as a supplementary method to conventional blood and biopsy culture testing, especially in culture-negative IE cases that are negative for IE by culture.

Keyword

16S rRNA gene; Infective endocarditis; Utility; PCR; Sequencing

MeSH Terms

Bacteria
Biopsy
Diagnosis*
DNA, Ribosomal*
Endocarditis*
Heart Valves
Humans
Medical Records
Methods
Polymerase Chain Reaction*
Streptococcus mitis
Thoracic Surgery
DNA, Ribosomal

Figure

  • Fig. 1 Comparison of blood culture, heart valve or vegetation biopsy culture, and/or 16S rDNA PCR results in cases treated for IE. The case numbers indicate positive results by blood culture, biopsy culture, and 16S rDNA PCR results (n=49, 20, and 43, respectively). Twenty cases were negative by all three methods.


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