A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus

  • Yihong Hu Pathogen Discovery and Big Data Center, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China
  • Zhenzhou Wan Medical Laboratory of Taizhou Fourth People’s Hospital, Taizhou, China
  • Yonglin Mu Key Laboratory of Computational Biology, Chinese Academy of Sciences-Max Planck Partner Institute for Computational Biology, Chinese Academy of Sciences, Shanghai, China
  • Yi Zhou Pathogen Discovery and Big Data Center, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China
  • Jia Liu Pathogen Discovery and Big Data Center, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China
  • Ke Lan State Key Laboratory of Virology, College of Life Sciences, Medical Research Institute, Wuhan University, Wuhan, China
  • Chiyu Zhang Pathogen Discovery and Big Data Center, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China
Keywords: RT-LAMP, RSV, SYTO 9, Sensitivity, Specificity, Clinical diagnosis

Abstract

Introduction: Human respiratory syncytial virus (hRSV) is a common respiratory virus closely related to respiratory tract infection (RTI). Rapid and accurate detection of hRSV is urgently needed to reduce the high morbidity and mortality due to hRSV infection.

Methodology: Here, we established a highly sensitive and specific reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for the rapid detection of A and B group hRSV simultaneously. The specific primer sets for hRSV A and B groups were designed in the M and M2-2 gene, respectively. SYTO 9 was used as the fluorescent dye for real-time monitoring of the amplification of hRSV RNA without cross reaction between hRSV A and B.

Results: The limit of detection (LOD) of our new method was 281.17 50% tissue culture infective doses (TCID50)/mL for hRSV A and 1.58 TCID50/mL for hRSV B. Using 90 clinical samples, a comparison to traditional RT-PCR was performed to validate this assay. The positivity rate of RT-LAMP and RT-PCR were 67.8% and 55.6%, respectively, and the positivity rate of RT-LAMP was significantly higher than RT-PCR (χ2 test, P < 0.01).

Conclusions: Compared with traditional RT-PCR method, the newly developed fluorescent RT-LAMP combined with well-designed primers and SYTO 9 is quite sensitive, specific, rapid and well applicable to hRSV clinical diagnosis.

Author Biography

Yihong Hu, Pathogen Discovery and Big Data Center, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China

Associate Professor

Published
2019-12-31
How to Cite
1.
Hu Y, Wan Z, Mu Y, Zhou Y, Liu J, Lan K, Zhang C (2019) A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus. J Infect Dev Ctries 13:1135-1141. doi: 10.3855/jidc.11549
Section
Original Articles