Direct detection of Corynebacterium striatum, Corynebacterium propinquum, and Corynebacterium simulans in sputum samples by high-resolution melt curve analysis
| Autor: | Xingzhao Ji, Haijian Zhou, Shuai Xu, Dan Li, Jingshan Zhang, Dongke Chen, Ming-Hui Li, Xiaotong Qiu, Lichao Han, Xuexin Hou, Mengtong Li, Zhenjun Li, Xuebing Wang, Lina Sun |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Genotyping Techniques 030106 microbiology Corynebacterium Biology Real-Time Polymerase Chain Reaction Melting curve analysis High Resolution Melt lcsh:Infectious and parasitic diseases High-resolution melt curve analysis 03 medical and health sciences Bacterial Proteins Limit of Detection Genetic variation medicine Humans lcsh:RC109-216 Bacterial identification Corynebacterium simulans Diagnostics DNA Primers Detection limit Corynebacterium propinquum Corynebacterium Infections Sputum Molecular biology Corynebacterium species 030104 developmental biology Infectious Diseases Parasitology medicine.symptom Research Article |
| Zdroj: | BMC Infectious Diseases BMC Infectious Diseases, Vol 21, Iss 1, Pp 1-9 (2021) |
| ISSN: | 1471-2334 |
| DOI: | 10.1186/s12879-020-05633-z |
| Popis: | Background Pulmonary infections caused by non-diphtheriae corynebacteria are increasing. However, rapid identification of Corynebacterium species poses a challenge due to the low genetic variation within the genus. Methods Three reference strains and 99 clinical isolates were used in this study. A qPCR followed by high-resolution melting (HRM) targeting ssrA was performed to simultaneously identify C. striatum, C. propinquum and C. simulans. To further evaluate this assay’s performance, 88 clinical sputum samples were tested by HRM and the detection results were compared with those of the traditional culture method and multiple cross-displacement amplification (MCDA) assay. Results The melting curve produced by a pair of universal primers generated species-specific HRM curve profiles and could distinguish the three target species from other related bacteria. The limit of detection of HRM assay for DNA from the three purified Corynebacterium species was 100 fg. Compared with the culture method, HRM detected 22 additional positive specimens, representing a 23.9% relative increase in detection rate. The HRM assay had 98.4% (95% confidence interval [CI], 90.5–99.9%) sensitivity and 100% (95% CI, 82.8–100%) specificity. Additionally, 95.5% concordance between HRM and MCDA (κ = 0.89 [95% CI, 0.79–0.99]) was noted. Conclusions The HRM assay was a simple, rapid, sensitive, and specific diagnostic tool for detecting C. striatum, C. propinquum, and C. simulans, with the potential to contribute to early diagnosis, epidemiological surveillance, and rapid response to outbreak. |
| Databáze: | OpenAIRE |
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