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https://doi.org/10.1016/j.anaerobe.2005.01.007

has been cited by the following article:

• TITLE: One Step Multiplex PCR for Identifications at Subspecies Level of Fusobacterium nucleatum and Fusobacterium necrophorum

  AUTHORS: Koji Umezawa, Sachiyo Hayashi, Osamu Tsuzukibashi, Akira Fukatsu, Satoshi Uchibori, Mana Fuchigami, Chiaki Komine, Takashi Asano, Masanobu Wakami, Taira Kobayashi, Masahiko Fukumoto, Takato Nomoto

  KEYWORDS: Fusobacterium nucleatum, Fusobacterium necrophorum, One-Step Multiplex PCR

  JOURNAL NAME: Open Journal of Stomatology, Vol.12 No.6, June 30, 2022

  ABSTRACT: Purpose: Fusobacterium nucleatum is an opportunistic pathogen involved in periodontal diseases, extraoral infections, and colorectal cancer. Fusobacterium necrophorum causes a variety of necrotic infections. F. nucleatum and F. necrophorum are classified into five and two subspecies, respectively. Conventional identification methods were technically hard to distinguish each subspecies of two Fusobacterium species accurately. The purpose of the present study was to design primers to identify two medically important Fusobacterium species at the subspecies level, using one-step multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the 16S ribosomal RNA (16S rRNA) gene, RNA polymerase B (rpoB) gene, and DNA gyrase subunit B (gyrB) of each subspecies of F. nucleatum and F. necrophorum. Results: These primers were able to distinguish each subspecies of F. nucleatum and F. necrophorum and did not display cross-reactivity with representative Fusobacterium species other than F. nucleatum and F. necrophorum. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and worked without requiring DNA extraction.