論文 - 詳細
| RRC ID | 47606 |
|---|---|
| 著者 | Ogawa M, Sugita S, Shimizu N, Watanabe K, Nakagawa I, Mochizuki M. |
| タイトル | Broad-range real-time PCR assay for detection of bacterial DNA in ocular samples from infectious endophthalmitis. |
| ジャーナル | Jpn J Ophthalmol |
| Abstract |
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 ≥ 10(7) copies/mL. Extraction with the Bacteria card(®) with lysozyme resulted in detection of all the tested strains at concentrations ≥ 10(6) 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 1-10 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. |
| 巻・号 | 56(6) |
| ページ | 529-35 |
| 公開日 | 2012-11-1 |
| DOI | 10.1007/s10384-012-0174-z |
| PMID | 22936171 |
| MeSH | Colony Count, Microbial DNA, Bacterial / analysis* Endophthalmitis / microbiology* Eye Infections, Bacterial / microbiology* Gram-Negative Bacteria / genetics Gram-Negative Bacteria / isolation & purification* Gram-Positive Bacteria / genetics Gram-Positive Bacteria / isolation & purification* Humans RNA, Bacterial / genetics RNA, Ribosomal, 16S / genetics* Real-Time Polymerase Chain Reaction / methods* Sensitivity and Specificity |
| IF | 1.725 |
| 引用数 | 17 |
| WOS 分野 | OPHTHALMOLOGY |
| リソース情報 | |
| 一般微生物 | JCM 8702 JCM 2414 JCM 5708 JCM 20313 JCM 1310 JCM 20266 JCM 1290 JCM 6425 JCM 20135 JCM 1662 JCM 6425 JCM 20914 |