A protocol for polymerase chain reaction detection of Enterococcus faecalis and Enterococcus faecium from the root canal

被引:25
作者
Molander, A
Lundquist, P
Papapanou, PN
Dahlén, G
Reit, C
机构
[1] Univ Gothenburg, Fac Odontol, Dept Endodontol Oral Diag, SE-40530 Gothenburg, Sweden
[2] Univ Gothenburg, Fac Odontol, Dept Oral Biochem, SE-40530 Gothenburg, Sweden
[3] Univ Gothenburg, Fac Odontol, Dept Oral Microbiol, SE-40530 Gothenburg, Sweden
关键词
enterococci; microbiology; PCR; root canal therapy;
D O I
10.1046/j.1365-2591.2002.00476.x
中图分类号
R78 [口腔科学];
学科分类号
1003 ;
摘要
Aim The present study was set up to develop a protocol for detection of Enterococcus faecalis and Enterococcus faecium from the root canal. Methodology A collection of type strains and clinical isolates of E. faecalis and E. faecium was used. Specific polymerase chain reaction (PCR) primers targeted against the 16S/23S rDNA intergenic region were used and PCR reactions were set up. PCR products were run on TBE-agarose gel and analysed. The sensitivity of the PCR system was studied using serial dilutions of (i) bacterial DNA and (ii) bacterial cells from E. faecalis. The specificity of the identification was tested against closely related species. Results All strains of E. faecalis and E. faecium produced identical amplicon profiles composed of two major bands corresponding to sizes of 320 and 420 bp. When amplifying DNA of higher purity, a third band of 600 bp became evident as well. Closely related species demonstrated single bands of various sizes and were easily distinguished from enterococci. The detection level of DNA from serial dilutions of DNA was 10(-13) g. The DNA extraction protocol from bacterial Cell Suspensions resulted in a detection level of 10 bacterial cells per sample. Conclusions The present study demonstrated a good potential for using PCR technology in the detection of E. faecalis and E. faecium from root canal samples. With a high specificity the methodology was able to detect 10 cells of F. faecalis.
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页码:1 / 6
页数:6
相关论文
共 30 条
[1]   Polymerase chain reaction detection of 8 putative periodontal pathogens in subgingival plaque of gingivitis and advanced periodontitis lesions [J].
Ashimoto, A ;
Chen, C ;
Bakker, I ;
Slots, J .
ORAL MICROBIOLOGY AND IMMUNOLOGY, 1996, 11 (04) :266-273
[2]  
Barry T, 1991, PCR Methods Appl, V1, P149
[3]  
BENDER I B, 1964, Oral Surg Oral Med Oral Pathol, V18, P527, DOI 10.1016/0030-4220(64)90403-7
[4]   Identification and antimicrobial susceptibility of enterococci isolated from the root canal [J].
Dahlén, G ;
Samuelsson, W ;
Molander, A ;
Reit, C .
ORAL MICROBIOLOGY AND IMMUNOLOGY, 2000, 15 (05) :309-312
[5]   A COMPARISON OF 2 TRANSPORT MEDIA FOR SALIVA AND SUBGINGIVAL SAMPLES [J].
DAHLEN, G ;
PIPATTANAGOVIT, P ;
ROSLING, B ;
MOLLER, AJR .
ORAL MICROBIOLOGY AND IMMUNOLOGY, 1993, 8 (06) :375-382
[6]   DETECTION OF GLYCOPEPTIDE RESISTANCE GENOTYPES AND IDENTIFICATION TO THE SPECIES LEVEL OF CLINICALLY RELEVANT ENTEROCOCCI BY PCR [J].
DUTKAMALEN, S ;
EVERS, S ;
COURVALIN, P .
JOURNAL OF CLINICAL MICROBIOLOGY, 1995, 33 (01) :24-27
[7]  
ENGSTROM B, 1964, ODONTOL TIDSKR, V72, P249
[8]   Variations in the susceptibilities of components of the endodontic microflora to biomechanical procedures [J].
Gomes, BPFA ;
Lilley, JD ;
Drucker, DB .
INTERNATIONAL ENDODONTIC JOURNAL, 1996, 29 (04) :235-241
[9]  
Hardie J.M., 1986, Bergey's Manual of Systematic Bacteriology, V2, P1068
[10]   In vitro susceptibility studies and detection of vancomycin resistance genes in clinical isolates of enterococci in Nagasaki, Japan [J].
Hirakata, Y ;
Yamaguchi, T ;
Izumikawa, K ;
Matsuda, J ;
Tomono, K ;
Kaku, M ;
Koga, H ;
Yamada, Y ;
Kohno, S ;
Kamihira, S .
EPIDEMIOLOGY AND INFECTION, 1997, 119 (02) :175-181