Zhihao Zhuo1,2,3, Jin Wang1,2,4, Wendi Chen1,2,4, Xiaosong Su1,2,4, Mengyuan Chen1,2,4, Mujin Fang1,2,4, Shuizhen He5, Shiyin Zhang6,7,8, Shengxiang Ge9,10,11, Jun Zhang1,2,4, Ningshao Xia1,2,4. 1. National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiang'an Campus of Xiamen University, Xiang'an South Road, Xiang'an District, Xiamen, Fujian, China. 2. State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Xiamen University, Xiang'an Campus of Xiamen University, South Xiang'an Rd, Xiamen, China. 3. School of Life Sciences, Xiamen University, Xiang'an Campus of Xiamen University, South Xiang'an Rd, Xiamen, China. 4. School of Public Health, Xiamen University, Xiang'an Campus of Xiamen University, South Xiang'an Rd, Xiamen, China. 5. Xiamen Center for Disease Control and Prevention, Shengguang Rd., Jimei District, Xiamen, China. 6. National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiang'an Campus of Xiamen University, Xiang'an South Road, Xiang'an District, Xiamen, Fujian, China. zhangshiyin@xmu.edu.cn. 7. State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Xiamen University, Xiang'an Campus of Xiamen University, South Xiang'an Rd, Xiamen, China. zhangshiyin@xmu.edu.cn. 8. School of Public Health, Xiamen University, Xiang'an Campus of Xiamen University, South Xiang'an Rd, Xiamen, China. zhangshiyin@xmu.edu.cn. 9. National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiang'an Campus of Xiamen University, Xiang'an South Road, Xiang'an District, Xiamen, Fujian, China. sxge@xmu.edu.cn. 10. State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Xiamen University, Xiang'an Campus of Xiamen University, South Xiang'an Rd, Xiamen, China. sxge@xmu.edu.cn. 11. School of Public Health, Xiamen University, Xiang'an Campus of Xiamen University, South Xiang'an Rd, Xiamen, China. sxge@xmu.edu.cn.
Abstract
BACKGROUND: Morbidity and mortality from influenza A (Flu A) have increased in recent years. Timely diagnosis and management are critical for disease control. Therefore, the development of a rapid, accurate, and portable analytical method for on-site analysis is imperative. OBJECTIVES: The aim of this work was to develop a rapid, on-site, automated assay for the detection of Flu A and to evaluate the assay. METHODS: A handheld instrument (TD-01) based on capillary convective polymerase chain reaction (PCR) was developed for rapid on-site detection of Flu A. Since a previous version of the instrument, an automated motion mechanism has been introduced to TD-01 to achieve RNA automated testing. The primers and probe used for Flu A detection were designed according to the Flu A gene sequence of matrix proteins. Finally, we evaluated the detection spectra, sensitivity, specificity, and diagnostic performance of the assay. RESULTS: The TD-01 was able to successfully automatically detect Flu A RNA within 30 min. Results for serially diluted viruses indicated that the lower limit of detection for Flu A was 0.1 TCID50/ml (50% tissue culture infective dose). After evaluating known virus stocks, including 15 strains of Flu A, four strains of Flu B, and two strains of respiratory syncytial virus (RSV), the assay had a favorable detection spectrum and no obvious cross-reactivity. Method verification based on 554 clinical samples indicated that the sensitivity and specificity of TD-01 were 98.30% (231/235) and 98.75% (315/319), respectively. CONCLUSIONS: The results indicate that Flu A detection by TD-01 is particularly suitable for on-site testing and has the potential for application in point-of-care testing.
BACKGROUND: Morbidity and mortality from influenza A (Flu A) have increased in recent years. Timely diagnosis and management are critical for disease control. Therefore, the development of a rapid, accurate, and portable analytical method for on-site analysis is imperative. OBJECTIVES: The aim of this work was to develop a rapid, on-site, automated assay for the detection of Flu A and to evaluate the assay. METHODS: A handheld instrument (TD-01) based on capillary convective polymerase chain reaction (PCR) was developed for rapid on-site detection of Flu A. Since a previous version of the instrument, an automated motion mechanism has been introduced to TD-01 to achieve RNA automated testing. The primers and probe used for Flu A detection were designed according to the Flu A gene sequence of matrix proteins. Finally, we evaluated the detection spectra, sensitivity, specificity, and diagnostic performance of the assay. RESULTS: The TD-01 was able to successfully automatically detect Flu A RNA within 30 min. Results for serially diluted viruses indicated that the lower limit of detection for Flu A was 0.1 TCID50/ml (50% tissue culture infective dose). After evaluating known virus stocks, including 15 strains of Flu A, four strains of Flu B, and two strains of respiratory syncytial virus (RSV), the assay had a favorable detection spectrum and no obvious cross-reactivity. Method verification based on 554 clinical samples indicated that the sensitivity and specificity of TD-01 were 98.30% (231/235) and 98.75% (315/319), respectively. CONCLUSIONS: The results indicate that Flu A detection by TD-01 is particularly suitable for on-site testing and has the potential for application in point-of-care testing.
Authors: James Mahony; Sylvia Chong; David Bulir; Alexandra Ruyter; Ken Mwawasi; Daniel Waltho Journal: J Clin Virol Date: 2013-07-01 Impact factor: 3.168
Authors: Marion Koopmans; Berry Wilbrink; Marina Conyn; Gerard Natrop; Hans van der Nat; Harry Vennema; Adam Meijer; Jim van Steenbergen; Ron Fouchier; Albert Osterhaus; Arnold Bosman Journal: Lancet Date: 2004-02-21 Impact factor: 79.321
Authors: Philipp P Nelson; Barbara A Rath; Paraskevi C Fragkou; Emmanouil Antalis; Sotirios Tsiodras; Chrysanthi Skevaki Journal: Front Cell Infect Microbiol Date: 2020-04-29 Impact factor: 5.293