Development of strain-specific PCR primers based on a DNA probe Fu12 for the identification of fusobacterium nucleatum subsp. nucleatum ATCC 25586T.
The objective of this study was to assess the strain-specificity of a DNA probe, Fu12, for Fusobacterium nucleatum subsp. nucleatum ATCC 25586T (F. nucleatum ATCC 25586T), and to develop sets of strain-specific polymerase chain reaction (PCR) primers. Strain-specificity was tested against 16 strains of F. nucleatum and 3 strains of distinct Fusobacterium species. Southern blot hybridization revealed that the Fu12 reacted exclusively with the HindIII-digested genomic DNA of F. nucleatum ATCC 25586T. The results of PCR revealed that three pairs of PCR primers, based on the nucleotide sequence of Fu12, generated the strain-specific amplicons from F. nucleatum ATCC 25586T. These results suggest that the DNA probe Fu12 and the three pairs of PCR primers could be useful in the identification of F. nucleatum ATCC 25586T, especially with regard to the determination of the authenticity of the strain.
Kim
Hwa-Sook
HS
Department of Oral Biochemistry, College of Dentistry, Chosun University, 375 Seo-Suk Dong, Dong-ku, Gwang-ju 501-759, Republic of Korea.
Song
Soo Keun
SK
Yoo
So Young
SY
Jin
Dong Chun
DC
Shin
Hwan Seon
HS
Lim
Chae Kwang
CK
Kim
Myung-Soo
MS
Kim
Jin-Soo
JS
Choe
Son-Jin
SJ
Kook
Joong-Ki
JK
D015139:Blotting, Southern; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005674:Fusobacterium Infections / Q000382:microbiology; D016967:Fusobacterium nucleatum / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008163:Luminescent Measurements; D010518:Periodontitis / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D015342:DNA Probes; D004269:DNA, Bacterial