A broad-range PCR technique for the diagnosis of infective endocarditis

Abstract

Infective endocarditis (IE) remains a severe and potentially fatal disease demanding sophisticated diagnostic strategies for detection of the causative microorganisms. The aim of the present study was to develop a broad-range 16S ribosomal RNA gene polymerase chain reaction in the routine diagnostic of IE for the early diagnosis of fatal disease. A broad-range PCR technique was selected and evaluated in terms of its efficiency in the diagnosis of endocarditis using 19 heart valves from patients undergoing cardiovascular surgeries at the Habib Bourguiba Hospital of Sfax, Tunisia, on the grounds of suspected IE. The results demonstrated the efficiency of this technique particularly in cases involving a limited number of bacteria since it helped to increase detection sensitivity. The technique proved to be efficient, particularly, in the bacteriological diagnosis of IE in contexts involving negative results from conventional culture methods and other contexts involving bacterial species that were not amenable to identification by phenotypic investigations. Indeed, the sequencing of the partial 16S ribosomal RNA gene revealed the presence of Bartonella henselae, Enterobacter sp., and Streptococcus pyogenes in three heart valves with the negative culture. It should be noted that the results obtained from the polymerase chain reaction-sequencing identification applied to the heart valve and the strain isolated from the same tissue were not consistent with the ones found by the conventional microbiological methods in the case of IE caused by Gemella morbillorum. In fact, the results from the molecular identification revealed the presence of Lactobacillus jensenii. Overall, the results have revealed that the proposed method is sensitive, reliable and might open promising opportunities for the early diagnosis of IE.

Infective Endocarditis Causing Native and Prosthetic Heart Valve Dysfunction

“…В связи с этим следует рассматривать метод ПЦР как необходимый диагностический анализ, а не только как дополняющий анализ при отрицательном результате микробиологического посева. Это связано с возможностью ПЦР определять следовые количества микробных клеток, в том числе не проявляющих жизнеспособность [76,77]. Однако стоит заметить, что ПЦР-диагностика является высокочувствительным методом, позволяющим захватывать даже случайные контаминанты из окружающей среды, вследствие чего стоит соблюдать особые требования к персоналу, помещениям и зонам проведения анализа.…”

Infektiöse Endokarditis – Update

“…Ähnlich der Therapiedauer bleiben die Ergebnisse entsprechender randomisierter Studien Voraussetzung zur gesicherten oralen Antibiotikatherapie. Die wesentlichen Herausforderungen einer erfolgreichen Antibiotikatherapie sind eine zunehmende bakterielle Toleranz und Resistenzentwicklung 16 .…”

Full pathogen characterisation: species identification including the detection of virulence factors and antibiotic resistance genes via multiplex DNA-assays

“…The currently most popular and clinically used tests are based on real-time polymerase chain reactions (PCRs) due to their relatively good sensitivity, specificity and speed 19 , 20 . Using highly conserved ribosomal RNA (rRNA) genes as targets allows usually a higher sensitivity since multiple copies of this genes are present in the genome 21 , 22 . The application as well as the variability of PCR are multitudinous, offering RT-PCR, isothermal PCR, loop-mediated isothermal amplification, or the recombinase polymerase amplification (RPA).…”