Broad-range real-time PCR assay for detection of bacterial DNA in ocular samples from infectious endophthalmitis

详细信息    查看全文

• 作者：Manabu Ogawa (1)
  Sunao Sugita (1) (2)
  Norio Shimizu (3)
  Ken Watanabe (3)
  Ichiro Nakagawa (4)
  Manabu Mochizuki (1)
  
• 关键词：Endophthalmitis ; Bacteria ; Polymerase chain reaction
• 刊名：Japanese Journal of Ophthalmology
• 出版年：2012
• 出版时间：November 2012
• 年：2012
• 卷：56
• 期：6
• 页码：529-535
• 全文大小：270KB
• 参考文献：1. Brun-Buisson C, Fartoukh M, Lechapt E, Honoré S, Zahar JR, Cerf C, et al. Contribution of blinded, protected quantitative specimens to the diagnostic and therapeutic management of ventilator-associated pneumonia. Chest. 2005;128:533-4. CrossRef
  2. Therese KL, Anand AR, Madhavan HN. Polymerase chain reaction in the diagnosis of bacterial endophthalmitis. Br J Ophthalmol. 1998;82:1078-2. CrossRef
  3. Chiquet C, Cornut PL, Benito Y, Thuret G, Maurin M, Lafontaine PO, et al. Eubacterial PCR for bacterial detection and identification in 100 acute postcataract surgery endophthalmitis. Invest Ophthalmol Vis Sci. 2008;49:1971-. CrossRef
  4. Sugita S, Shimizu N, Watanabe K, Katayama M, Horie S, Ogawa M, et al. Diagnosis of bacterial endophthalmitis by broad-range quantitative PCR. Br J Ophthalmol. 2011;95:345-. CrossRef
  5. Takai K, Horikoshi K. Rapid detection and quantification of members of the archaeal community by quantitative PCR using fluorogenic probes. Appl Environ Microbiol. 2000;66:5066-2. CrossRef
  6. Zucol F, Ammann RA, Berger C, Aebi C, Altwegg M, Niggli FK, et al. Real-time quantitative broad-range PCR assay for detection of the 16S rRNA gene followed by sequencing for species identification. J Clin Microbiol. 2006;44:2750-. CrossRef
  7. Usui N, Uno T, Oki K, Oshika T, Ohashi Y, Ogura Y, et al. Nationwide surveillance of postoperative endophthalmitis related to cataract surgery. Jpn J Ophthalmic Surg. 2006;19:73-.
  8. Aaberg TM Jr, Flynn HW Jr, Schiffman J, Newton J. Nosocomial acute-onset postoperative endophthalmitis survey. A 10-year review of incidence and outcomes. Ophthalmology. 1998;105:1004-0. CrossRef
  9. Vafidis G. Propionibacterium acnes endophthalmitis. Br J Ophthalmol. 1991;75:706. CrossRef
  10. Boldt HC, Pulido JS, Blodi CF, Folk JC. Rural endophthalmitis. Ophthalmology. 1989;96:1722-.
  11. Jackson TL, Eykyn SJ, Graham EM, Stanford MR. Endogenous bacterial endophthalmitis: a 17-year prospective series and review of 267 reported cases. Surv Ophthalmol. 2003;48:403-3. CrossRef
  12. Chen YJ, Kuo HK, Wu PC, Kuo ML, Tsai HH, Liu CC, et al. A 10-year comparison of endogenous endophthalmitis outcomes: an east Asian experience with / Klebsiella pneumoniae infection. Retina. 2004;24:383-0. CrossRef
  13. Callegan MC, Engelbert M, Parke DW 2nd, Jett BD, Gilmore MS. Bacterial endophthalmitis: epidemiology, therapeutics, and bacterium–host interactions. Clin Microbiol Rev. 2002;15:111-4. CrossRef
  14. Callegan MC, Gilmore MS, Gregory M, Ramadan RT, Wiskur BJ, Moyer AL, et al. Bacterial endophthalmitis: therapeutic challenges and host–pathogen interactions. Prog Retin Eye Res. 2007;26:189-03. CrossRef
  15. Tomaso H, Kattar M, Eickhoff M, Wernery U, Al Dahouk S, Straube E, et al. Comparison of commercial DNA preparation kits for the detection of Brucellae in tissue using quantitative real-time PCR. BMC Infect Dis. 2010;10:100. CrossRef
  16. Tsai YL, Olson BH. Rapid method for direct extraction of DNA from soil and sediments. Appl Environ Microbiol. 1991;57:1070-.
  17. Lee YK, Kim HW, Liu CL, Lee HK. A simple method for DNA extraction from marine bacteria that produce extracellular materials. J Microbiol Methods. 2003;52:245-0. CrossRef
  18. Callewaert L, Aertsen A, Deckers D, Vanoirbeek KG, Vanderkelen L, Van Herreweghe JM, et al. A new family of lysozyme inhibitors contributing to lysozyme tolerance in gram-negative bacteria. PLoS Pathog. 2008;4:e1000019. doi:10.1371/journal.ppat.1000019 . CrossRef
  19. Davis KM, Akinbi HT, Standish AJ, Weiser JN. Resistance to mucosal lysozyme compensates for the fitness deficit of peptidoglycan modifications by / Streptococcus pneumoniae. PLoS Pathog. 2008;4:e1000241. doi:10.1371/journal.ppat.1000241 . CrossRef
  20. Vollmer T, St?rmer M, Kleesiek K, Dreier J. Evaluation of novel broad-range real-time PCR assay for rapid detection of human pathogenic fungi in various clinical specimens. J Clin Microbiol. 2008;46:1919-6. CrossRef
  
• 作者单位：Manabu Ogawa (1)
  Sunao Sugita (1) (2)
  Norio Shimizu (3)
  Ken Watanabe (3)
  Ichiro Nakagawa (4)
  Manabu Mochizuki (1)
  
  1. Department of Ophthalmology and Visual Science, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
  2. Laboratory for Retinal Regeneration, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, 650-0047, Japan
  3. Division of Medical Science, Department of Virology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
  4. Department of Section of Bacterial Pathogenesis, Graduate School of Medicine and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
  
• ISSN：1613-2246

文摘

Background To evaluate a broad-range real-time polymerase chain reaction (PCR) targeting the bacterial 16S rRNA gene for detection of bacterial DNA in infectious endophthalmitis. Methods The bacterial 16S rRNA gene was measured by quantitative real-time PCR. For the assay, bacterial DNA was prepared from 12 Gram-positive and 4 Gram-negative strains. To determine the optimum method for DNA extraction, four extraction procedures were selected by using DNA extraction program cards with and without the use of lysozyme. To evaluate PCR sensitivity, PCR fragments were amplified from Staphylococcus aureus and Escherichia coli DNA. Results DNA extraction using the Bacteria card? without enzymes resulted in detection of all the tested strains at concentrations ?07?copies/mL. Extraction with the Bacteria card? with lysozyme resulted in detection of all the tested strains at concentrations ?06?copies/mL, indicative of no significant difference between the two procedures. DNA extraction using the Virus card?, both with and without enzymes, resulted in reduced efficiency of detection of all strains compared with use of the Bacteria card?. The PCR could detect as few as 1-0?colony-forming units (CFU) in diluted vitreous samples per reaction, and all tested bacterial species known to cause endophthalmitis were detected. Conclusions Bacterial 16S-specific PCR can comprehensively detect the main causative bacteria of clinically suspected endophthalmitis.