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Research Journal of Microbiology
  Year: 2018 | Volume: 13 | Issue: 2 | Page No.: 93-99
 
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Identification of HACEK Group Bacteria from Blood Samples of Patients with Infective Endocarditis by PCR-RFLP of the 16s rRNA Gene

Taichi Ishikawa, Yu Shimoyama, Yoshitoyo Kodama, Shihoko Tajika, Shigenobu Kimura and Minoru Sasaki

Abstract:
Background and Objective: Identification of specific HACEK bacteria that inhabit the human oral cavity and cause infective endocarditis (IE) is difficult because conventional culture methods produce inconclusive results in cases of fastidious and slow-growing organisms. Although the study of a rapid and sensitive identification method for identification of HACEK bacteria based on polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis of the 16S rRNA gene have been reported, the procedures were not revealed. Herein, in this study, the detail of the method conditions and procedures were described and assessed its usefulness in analyzing clinical samples using eight clinical isolates from patients with IE. Materials and Methods: The bacteria were analyzed by the 16S rRNA gene PCR-RFLP method using HinfI and MspI. The isolate from patients were further subjected to species-specific identification with biochemical identification kits. Results: Seven isolates were identified as Streptococcus intermedius (2×), Abiotrophia defective (2×), Granulicatella adiacR-RFLP results of the other clinical isolates wereens, Streptococcus salivarius and Staphylococcus epidermidis using a biochemical identification kit (eighth was unidentified). The HACEK bacteria and the isolates were further subjected to PCR-RFLP analysis of the 16S rRNA gene. Typical restriction patterns were obtained by combination digestion with HinfI and MspI. The patterns of the unidentified isolate were same as those of C. hominis, thereby confirming the identification of the causative pathogen. The PC identical to those with the identification kits. Conclusion: The PCR-RFLP analysis of the 16S rRNA gene is applicable for definitive diagnosis of HACEK group, fastidious growing bacteria and in samples with unidentifiable bacteria using conventional biochemical identification kits.
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How to cite this article:

Taichi Ishikawa, Yu Shimoyama, Yoshitoyo Kodama, Shihoko Tajika, Shigenobu Kimura and Minoru Sasaki, 2018. Identification of HACEK Group Bacteria from Blood Samples of Patients with Infective Endocarditis by PCR-RFLP of the 16s rRNA Gene. Research Journal of Microbiology, 13: 93-99.

URL: https://scialert.net/abstract/?doi=jm.2018.93.99

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