論文 - 詳細
| RRC ID | 71303 |
|---|---|
| 著者 | Sasano M, Seki M, Takano C, Komine-Aizawa S, Hayakawa S. |
| タイトル | An improved primer design for the loop-mediated isothermal amplification (LAMP) method to detect oxacillinase (OXA)-48 β-lactamase genes in Gram-negative bacteria for clinical applications. |
| ジャーナル | J Infect Chemother |
| Abstract |
INTRODUCTION:Recently, increased frequencies of carbapenemase-producing Enterobacteriaceae have been reported worldwide. Among multiple genetic subtypes, oxacillinase (OXA)-48 β-lactamase-producing strains have been associated with inbound infection because they have been detected predominantly in patients who traveled outside of Japan. However, a recent case report of OXA-48 β-lactamase-producing Enterobacteriaceae suggested the latent spread of domestic infections. Due to a lack of specific inhibitors, culture-based detection of OXA-48 β-lactamase-producing bacteria is difficult. Thus, DNA-based detection methods, including PCR, direct sequencing and loop-mediated isothermal amplification (LAMP), have been employed. Among these methods, LAMP detection is more favorable than other methods because of its technical simplicity and low cost. METHODS:We designed novel LAMP primers to detect OXA-48 β-lactamase-producing bacteria and investigated their possible clinical applications with bacterial genome-spiked human materials (cerebrospinal fluid, blood, feces, urine, and sputum). We evaluated the specificity of the LAMP primers using 37 bacterial strains: 8 standard, 9 reference, and 20 clinical Gram-negative strains. RESULTS:Our LAMP primers detected 10 copies of the OXA-48 type β-lactamase gene and exhibited no cross reactivity with other β-lactamase genes. Sensitivity was not influenced in any clinical sample, in contrast to PCR detection, which was strongly inhibited by substances in fecal samples. CONCLUSIONS:These results suggest the superior performance of LAMP compared with conventional PCR for detecting the OXA-48 type β-lactamase gene in various clinical samples. |
| 巻・号 | 27(7) |
| ページ | 1005-1012 |
| 公開日 | 2021-7-1 |
| DOI | 10.1016/j.jiac.2021.02.016 |
| PII | S1341-321X(21)00056-8 |
| PMID | 33814349 |
| MeSH | Bacterial Proteins / genetics Gram-Negative Bacteria / genetics Humans Japan Molecular Diagnostic Techniques Nucleic Acid Amplification Techniques* Sensitivity and Specificity beta-Lactamases* / genetics |
| リソース情報 | |
| 一般微生物 | JCM1232 JCM12093 JCM1669 JCM20624 |