A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus
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...
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The Journal of Infection in Developing Countries
2019-12-01
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| Series: | Journal of Infection in Developing Countries |
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| Online Access: | https://jidc.org/index.php/journal/article/view/11549 |
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| author | Yihong Hu Zhenzhou Wan Yonglin Mu Yi Zhou Jia Liu Ke Lan Chiyu Zhang |
| author_facet | Yihong Hu Zhenzhou Wan Yonglin Mu Yi Zhou Jia Liu Ke Lan Chiyu Zhang |
| author_sort | Yihong Hu |
| collection | DOAJ |
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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.
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| format | Article |
| id | doaj-art-dd4bedcec1914455a2d9493a337e0067 |
| institution | OA Journals |
| issn | 1972-2680 |
| language | English |
| publishDate | 2019-12-01 |
| publisher | The Journal of Infection in Developing Countries |
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| series | Journal of Infection in Developing Countries |
| spelling | doaj-art-dd4bedcec1914455a2d9493a337e00672025-08-20T02:27:18ZengThe Journal of Infection in Developing CountriesJournal of Infection in Developing Countries1972-26802019-12-01131210.3855/jidc.11549A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virusYihong Hu0Zhenzhou Wan1Yonglin Mu2Yi Zhou3Jia Liu4Ke Lan5Chiyu Zhang6Pathogen Discovery and Big Data Center, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, ChinaMedical Laboratory of Taizhou Fourth People’s Hospital, Taizhou, ChinaKey Laboratory of Computational Biology, Chinese Academy of Sciences-Max Planck Partner Institute for Computational Biology, Chinese Academy of Sciences, Shanghai, ChinaPathogen Discovery and Big Data Center, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, ChinaPathogen Discovery and Big Data Center, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, ChinaState Key Laboratory of Virology, College of Life Sciences, Medical Research Institute, Wuhan University, Wuhan, ChinaPathogen Discovery and Big Data Center, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China 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. https://jidc.org/index.php/journal/article/view/11549RT-LAMPRSVSYTO 9SensitivitySpecificityClinical diagnosis |
| spellingShingle | Yihong Hu Zhenzhou Wan Yonglin Mu Yi Zhou Jia Liu Ke Lan Chiyu Zhang A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus Journal of Infection in Developing Countries RT-LAMP RSV SYTO 9 Sensitivity Specificity Clinical diagnosis |
| title | A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus |
| title_full | A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus |
| title_fullStr | A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus |
| title_full_unstemmed | A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus |
| title_short | A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus |
| title_sort | quite sensitive fluorescent loop mediated isothermal amplification for rapid detection of respiratory syncytial virus |
| topic | RT-LAMP RSV SYTO 9 Sensitivity Specificity Clinical diagnosis |
| url | https://jidc.org/index.php/journal/article/view/11549 |
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