title: Comparative molecular and microbiological diagnosis of 19 infective endocarditis cases in which causative microbes were identified by PCR-based DNA sequencing from the excised heart valves.
abstract: Infective endocarditis (IE) is traditionally diagnosed by microbiological analysis of blood cultures, following which therapeutic antibiotics are chosen based on antimicrobial sensitivity tests. However, such conventional techniques do not always lead to an accurate etiological diagnosis. Recently, PCR analysis of the 16S rRNA gene has been employed to identify organisms isolated from excised heart valves. In this study, we analyzed 19 valve samples from patients with confirmed IE, as identified by Duke's criteria. Using broad-range PCR amplification, followed by direct gene sequencing, pathological agents were identified in all samples. Although blood cultures yielded negative results in 4 cases, PCR analysis of valve samples showed positive identification of causative organisms. In 3 cases, there was a difference between blood culture and PCR in identification of pathological agents, which are likely to be misidentified by the conventional method based on the phenotypic database. Postoperative antibiotics were chosen considering the severity of lesions and the results of PCR, Gram staining, and valve cultures. All patients were cured without relapse. The broad-range PCR method was therefore beneficial for the management of IE because it enabled us to identify pathogens directly from the site of infection, even organisms that were difficult to culture or likely to be misidentified by the conventional culture method. Identification of the agents provided precise knowledge of the microbiological spectrum involved in the cases of IE.
authors: Department of Infectious Diseases and Infection Control, Saitama International Medical Center, Saitama Medical University, 1397-1 Yamane, Hidaka, Saitama 350-1298, Japan. miyazato@saitama-med.ac.jp
authors: Akiko
authors: A
authors: Miyazato
authors: Kiyofumi
authors: K
authors: Ohkusu
authors: Mimiko
authors: M
authors: Tabata
authors: Kazuhiko
authors: K
authors: Uwabe
authors: Toru
authors: T
authors: Kawamura
authors: Yoshimi
authors: Y
authors: Tachi
authors: Takayuki
authors: T
authors: Ezaki
authors: Hiroshi
authors: H
authors: Niinami
authors: Kotaro
authors: K
authors: Mitsutake
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D016470:Bacteremia / Q000175:diagnosis / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry; D004697:Endocarditis, Bacterial / Q000097:blood / Q000175:diagnosis / Q000382:microbiology*; D005260:Female; D016908:Gram-Positive Bacterial Infections / Q000175:diagnosis / Q000382:microbiology; D006095:Gram-Positive Cocci / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006351:Heart Valves / Q000382:microbiology*; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial