The Journal of heart valve disease · 2013
16S rRNA sequencing as a diagnostic tool in the identification of culture-negative endocarditis in surgically treated patients.
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Kumar Naveen V, Menon Thangam, Pathipati Padmaja, Cherian Kotturathu Mammen
Аннотация
BACKGROUND AND AIM OF THE STUDY: Infective endocarditis (IE) is a worldwide problem, and at least one-third of cases are culture-negative despite the use of appropriate laboratory techniques.
METHODS: A broad-range polymerase chain reaction (PCR) amplification was performed of the 16S rRNA gene, followed by single-strand sequencing for 26 surgically removed heart valves from patients with culture-negative endocarditis who had undergone valve repair or replacement.
RESULTS: Two of the 26 patients were PCR-positive, and sequencing of the amplicon identified the etiological agent. Gram-stained smears of the heart valves were positive in both cases. Three of the remaining 24 cases which were negative by PCR also showed the presence of micro-organisms in Gram-stained smears.
CONCLUSION: The study results emphasize that, in suspected IE cases when there is no growth in culture, a combination of microscopy and 16S rRNA sequencing can be used to identify the pathogen in excised valvular tissue.
METHODS: A broad-range polymerase chain reaction (PCR) amplification was performed of the 16S rRNA gene, followed by single-strand sequencing for 26 surgically removed heart valves from patients with culture-negative endocarditis who had undergone valve repair or replacement.
RESULTS: Two of the 26 patients were PCR-positive, and sequencing of the amplicon identified the etiological agent. Gram-stained smears of the heart valves were positive in both cases. Three of the remaining 24 cases which were negative by PCR also showed the presence of micro-organisms in Gram-stained smears.
CONCLUSION: The study results emphasize that, in suspected IE cases when there is no growth in culture, a combination of microscopy and 16S rRNA sequencing can be used to identify the pathogen in excised valvular tissue.