1887

Abstract

Accurate identification of species remains a challenge due to the complexities of taxonomy and insufficient discriminatory power of traditional techniques. We report the development of a molecular technique that utilizes real-time PCR-based high-resolution melting (HRM) analysis for differentiation of the most common species. Based on a novel intergenic region sequence, , and were clearly distinguished from one another by HRM analysis. The limit of detection of the HRM assay for purified spp. DNA was at least 10 fg. No false positives were observed for specificity testing of 20 non-target clinical samples. In comparison to established matrix-assisted laser desorption/ionization-time of flight MS, the HRM assay improved the identification of . Additionally, all the products of PCR were verified by direct sequencing. In conclusion, the developed molecular assay allows simultaneous detection and differentiation of , and with high sensitivity and specificity.

Funding
This study was supported by the:
  • the China Special Grant for the Prevention and Control of Infectious Diseases (Award 2018ZX10734401;2018ZX10734404)
    • Principle Award Recipient: Xuexin Hou
  • the China Special Grant for the Prevention and Control of Infectious Diseases (Award 2017ZX10303401)
    • Principle Award Recipient: Zhenjun Li
  • National Key R&D Program of China (Award 2017YFC1200303)
    • Principle Award Recipient: Shuai Xu
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/content/journal/jmm/10.1099/jmm.0.001205
2020-06-01
2021-07-27
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