혈액 배양 액체배지에서 세균성 병원균 검출을 위한 16S rDNA PCR의평가
Evaluation of a 16S rDNA PCR Assay for Detection of Bacterial Pathogens in Blood Culture Broth
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Background: Rapid detection of pathogens in blood is important in patient management, because the mortality rate associated with bloodstream infections is very high. We evaluated the efficiency of a 16S rDNA PCR assay for the detection of various pathogens in blood culture broth in a clinical laboratory. Methods: 16S rDNA PCR was performed on 221 blood culture bottles consisting of 99 culturepositive and 122 culture-negative samples. The results were compared with conventional culture methods. We also compared the efficiency of three DNA extraction and purification methods using proteinase K, triton X-100, and benzyl alcohol-guanidine DNA extraction of blood culture broths. Results: The 16S rDNA PCR method detected 95 (12 Staphylococcus aureus, 27 coagulase negative staphylococci, 10 enterococci, 5 streptococci, 37 gram negative bacilli, 4 corynebacteria) of 99 positive culture bottles. Four false-negative results were obtained for bottles containing 2 Corynebacterium, 1 Escherichia coli, and 1 S. aureus species. All 122 bottles that showed no blood culture growth were negative by 16S rDNA PCR. Overall, the sensitivity, specificity, positive predictive values and negative predictive values of 16S rDNA PCR relative to the culture results were 96.0%, 100%, 100%, and 96.8%, respectively. Among the three DNA extraction methods, the benzyl alcohol-guanidine method was most effective. Conclusion: The 16S rDNA PCR assay is a rapid and efficient means of detecting various pathogens in the blood and has great potential for use in the clinical microbiology laboratory.
Abstract
Background: Rapid detection of pathogens in blood is important in patient management, because the mortality rate associated with bloodstream infections is very high. We evaluated the efficiency of a 16S rDNA PCR assay for the detection of various pathogens in blood culture broth in a clinical laboratory. Methods: 16S rDNA PCR was performed on 221 blood culture bottles consisting of 99 culturepositive and 122 culture-negative samples. The results were compared with conventional culture methods. We also compared the efficiency of three DNA extraction and purification methods using proteinase K, triton X-100, and benzyl alcohol-guanidine DNA extraction of blood culture broths. Results: The 16S rDNA PCR method detected 95 (12 Staphylococcus aureus, 27 coagulase negative staphylococci, 10 enterococci, 5 streptococci, 37 gram negative bacilli, 4 corynebacteria) of 99 positive culture bottles. Four false-negative results were obtained for bottles containing 2 Corynebacterium, 1 Escherichia coli, and 1 S. aureus species. All 122 bottles that showed no blood culture growth were negative by 16S rDNA PCR. Overall, the sensitivity, specificity, positive predictive values and negative predictive values of 16S rDNA PCR relative to the culture results were 96.0%, 100%, 100%, and 96.8%, respectively. Among the three DNA extraction methods, the benzyl alcohol-guanidine method was most effective. Conclusion: The 16S rDNA PCR assay is a rapid and efficient means of detecting various pathogens in the blood and has great potential for use in the clinical microbiology laboratory.
참고문헌 (15)
- "Detection of bacteraemia in critically ill patients using 16S rDNA polymerase chain reaction and DNA sequencing", 27, pp.1269~1273 (2001)
- "Detection of bacteraemia in patients with fever and neutropenia using 16S rRNA geneamplification by polymerase chain reaction", 17, pp.247~253 (1998)
- "Improved amplification of microbial DNA from blood cultures by removal of the PCR inhibitor sodium polyanetholesulfonate", 36, pp.2810~2816 (1998)
- "Rapid identification of Candida species in blood cultures by a clinically useful PCR method", 35, pp.1454~1459 (1997)
- "Chemotaxonomic and phylogenetic analyses of Sphigomonas strains isolated from ears of plants in the family Gramineae and a proposal of Sphingomonas roseoflava sp", 46, pp.9~18 (2000)
- "Use of PCR with universal primers and restriction endonuclease digestions for detection and identification of common bacterial pathogens in cerebrospinal fluid", 38, pp.2076~2080 (2000)
- "A simple and sensitive method to extract bacterial,yeast and fungal DNA from blood culture material", 42, pp.139~147 (2000)
- "Use of restriction fragment length polymorphism of the PCR-amplified 16S rRNA gene for the identification of Aeromonasspp", 38, pp.2023~2025 (2000)
- "Development of polymerase chain reaction primer sets for diagnosis of lyme disease and for species-specific identification of lyme disease isolates by 16S rRNA signature nucleotide analysis", 30, pp.2830~4 (1992)
- "Touchdown enzyme time release-PCR for detection and identification of Chlamydia trachomatis, C. pneumoniae, and C. psittaciusing the 16S and 16S-23S spacer rRNA genes", 38, pp.1085~1093 (2000)
- "Myoglobin as a polymerase chain reaction (PCR) inhibitor: a limitation for PCR from skeletal muscle tissue avoided by the use of Thermus thermophilus polymerase", 21, pp.1064~1067 (1998)
- "Purification and characterization of PCR - inhibitory components in blood cells", 39, pp.485~493 (2001)
- "Inhibition of PCR by aqueous and vitreous fluids", 33, pp.2643~2646 (1995)
- "Comparison of 16S rRNA gene PCR and BACTEC 9240 for detection of neonatal bacteremia", 38, pp.2574~2578 (2000)
- "Evaluation of an extended diagnostic PCR assay of detection and verification of the common cause of bacterial meningitis in CSF and other biological samples", 13, pp.49~60 (1999)