Journal of the korean academy of Pediatric Dentistry (대한소아치과학회지)

Korean Academy of Pediatric Dentistry (대한소아치과학회)
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Rapid Detection of Pathogens Associated with Dental Caries and Periodontitis by PCR Using a Modified DNA Extraction Method

PCR을 이용한 치아우식증 및 치주염 연관 병원체의 빠른 검출

  • Kim, Jaehwan (Department of Pediatric Dentistry and Institute of Oral Bioscience, School of Dentistry, Chonbuk National University) ;
  • Kim, Miah (Department of Pediatric Dentistry and Institute of Oral Bioscience, School of Dentistry, Chonbuk National University) ;
  • Lee, Daewoo (Department of Pediatric Dentistry and Institute of Oral Bioscience, School of Dentistry, Chonbuk National University) ;
  • Baik, Byeongju (Department of Pediatric Dentistry and Institute of Oral Bioscience, School of Dentistry, Chonbuk National University) ;
  • Yang, Yeonmi (Department of Pediatric Dentistry and Institute of Oral Bioscience, School of Dentistry, Chonbuk National University) ;
  • Kim, Jaegon (Department of Pediatric Dentistry and Institute of Oral Bioscience, School of Dentistry, Chonbuk National University)
  • 김재환 (전북대학교 치의학전문대학원 소아치과학교실 및 구강생체과학연구소) ;
  • 김미아 (전북대학교 치의학전문대학원 소아치과학교실 및 구강생체과학연구소) ;
  • 이대우 (전북대학교 치의학전문대학원 소아치과학교실 및 구강생체과학연구소) ;
  • 백병주 (전북대학교 치의학전문대학원 소아치과학교실 및 구강생체과학연구소) ;
  • 양연미 (전북대학교 치의학전문대학원 소아치과학교실 및 구강생체과학연구소) ;
  • 김재곤 (전북대학교 치의학전문대학원 소아치과학교실 및 구강생체과학연구소)
  • Received : 2014.06.16
  • Accepted : 2014.09.17
  • Published : 2014.11.30
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Abstract

DNA extraction is a prerequisite for the identification of pathogens in clinical samples. Commercial DNA extraction kits generally involve time-consuming and laborious multi-step procedures. In the present study, our modified DNA isolation method for saliva samples allows for the quick detection of pathogens associated with dental caries or periodontitis by PCR within 1 h. To release DNA from the bacteria, 1 min of boiling was adequate, and the resulting isolated DNA can be used many times and is suitable for long term storage of at least 13 months at $4^{\circ}C$, and even longer at $-20^{\circ}C$. In conclusion, our modified DNA extraction method is simple, rapid, and cost-effective, and suitable for preparing DNA from clinical samples for PCR for the rapid detection of oral pathogens from saliva.

구강 병원체의 검출 방법은 여러 가지가 있지만 그 중 PCR을 이용한 검출이 확실하고 빠른 방법으로 알려져 있다. PCR을 위한 많은 DNA 추출법이 사용되고 있으나 상업적인 DNA 추출 kit들은 일반적으로 가격이 비싸고 절차가 여러 단계로 되어있으며, 그 외의 방법은 페놀과 클로로포름과 같은 유해한 화학물질을 써야하는 등의 단점이 있다. 이 연구에서 NaOH 용액을 이용한 개선된 DNA 추출 방법은 치아우식증, 치주염과 관련된 병원체를 빠르고 간단하며 비용-효율적으로 검출하였다. 세균으로부터 DNA를 추출하기 위한 boiling은 기존의 10분이 아닌 1분으로 충분하였고 $4^{\circ}C$에서 최소 13개월 이상 DNA의 보관이 가능하였으며 sonication 유무에 따른 차이는 없었다. 따라서, 이 방법은 상업적인 kit나 유해한 화학물질을 쓰지 않고서도 타액 표본으로부터 직접적으로 빠른 시간 내에 DNA를 추출하여 병원체의 유무 결과를 확인하는데 매우 적합할 것으로 생각한다.

Keywords

References

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