Performance Evaluation of a Novel Ultrafast Molecular Diagnostic Device Integrated With Microfluidic Chips and Dual Temperature Modules
Ultrafast, portable, and inexpensive molecular diagnostic platforms are critical for clinical diagnosis and on-site detection. There are currently no available real-time polymerase chain reaction (PCR) devices able to meet the demands of point-of-care testing, as the heating and cooling processes ca...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.895236/full |
| _version_ | 1811257815065427968 |
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| author | Shan Lin Shan Lin Xiaojun Song Kun Zhu Quanyu Shao Yinhang Chen Wei Cheng Zhijing Lei Yu Chen Yu Chen Yun Luo Dazhi Jin Dazhi Jin Dazhi Jin |
| author_facet | Shan Lin Shan Lin Xiaojun Song Kun Zhu Quanyu Shao Yinhang Chen Wei Cheng Zhijing Lei Yu Chen Yu Chen Yun Luo Dazhi Jin Dazhi Jin Dazhi Jin |
| author_sort | Shan Lin |
| collection | DOAJ |
| description | Ultrafast, portable, and inexpensive molecular diagnostic platforms are critical for clinical diagnosis and on-site detection. There are currently no available real-time polymerase chain reaction (PCR) devices able to meet the demands of point-of-care testing, as the heating and cooling processes cannot be avoided. In this study, the dual temperature modules were first designed to process microfluidic chips automatically circulating between them. Thus, a novel ultrafast molecular diagnostic real-time PCR device (approximately 18 and 23 min for DNA and RNA detection, respectively) with two channels (FAM and Cy5) for the detection of 12 targets was developed. The device contained three core functional components, including temperature control, optics, and motion, which were integrated into a portable compact box. The temperature modules accurately control temperature in rapid thermal cycles with less than ±0.1 °C, ±1 °C and ±0.5 °C for the temperature fluctuation, uniformity, and error of indication, respectively. The average coefficient of variation (CV) of the fluorescence intensity (FI) for all 12 wells was 2.3% for FAM and 2.7% for Cy5. There was a good linear relationship between the concentrations of fluorescent dye and the FIs of FAM and Cy5(R2 = 0.9990 and 0.9937), and the average CVs of the Ct values calculated by the embedded software were 1.4% for FAM and Cy5, respectively. The 100 double-blind mocked sputum and 249 clinical stool samples were analyzed by the ultrafast real-time PCR device in comparison with the DAAN Gene SARS-CoV-2 kit run on the ABI 7500 instrument and Xpert C. difficile/Epi, respectively. Among the 249 stool samples, the ultrafast real-time PCR device detected toxigenic C. difficile in 54 samples (54/249, 21.7%) with a specificity and positive predictive values of 99.0 and 96.3%, which were higher than the Xpert C. difficile/Epi values of 94.4 and 88.1% (p > 0.05). The ultrafast real-time PCR device detected 15 SARS-CoV-2 positive samples, which has a 100% concordance with that obtained by the DAAN Gene SARS-CoV-2 kit. This study demonstrated that the ultrafast real-time PCR device integrated with microfluidic chips and dual temperature modules is an ultrafast, reliable, easy-to-use, and cost-effective molecular diagnostic platform for clinical diagnosis and on-site testing, especially in resource-limited settings. |
| first_indexed | 2024-04-12T18:04:28Z |
| format | Article |
| id | doaj.art-8547a110bfda4df58f203efb1b1b1a9d |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-12T18:04:28Z |
| publishDate | 2022-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-8547a110bfda4df58f203efb1b1b1a9d2022-12-22T03:22:03ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-05-011010.3389/fbioe.2022.895236895236Performance Evaluation of a Novel Ultrafast Molecular Diagnostic Device Integrated With Microfluidic Chips and Dual Temperature ModulesShan Lin0Shan Lin1Xiaojun Song2Kun Zhu3Quanyu Shao4Yinhang Chen5Wei Cheng6Zhijing Lei7Yu Chen8Yu Chen9Yun Luo10Dazhi Jin11Dazhi Jin12Dazhi Jin13School of Laboratory Medicine, Hangzhou Medical College, Hangzhou, ChinaKey Laboratory of Biomarkers and In Vitro Diagnosis Translation of Zhejiang Province, Hangzhou, ChinaDepartment of Clinical Laboratory, Laboratory Medicine Center, Zhejiang Provincial People’s Hospital, Hangzhou Medical College, Hangzhou, ChinaHangzhou Biochip for Diagnosis Technology Co., Ltd., Hangzhou, ChinaHangzhou Biochip for Diagnosis Technology Co., Ltd., Hangzhou, ChinaHangzhou Biochip for Diagnosis Technology Co., Ltd., Hangzhou, ChinaHangzhou Biochip for Diagnosis Technology Co., Ltd., Hangzhou, ChinaHangzhou Biochip for Diagnosis Technology Co., Ltd., Hangzhou, ChinaSchool of Laboratory Medicine, Hangzhou Medical College, Hangzhou, ChinaKey Laboratory of Biomarkers and In Vitro Diagnosis Translation of Zhejiang Province, Hangzhou, ChinaSchool of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, AustraliaSchool of Laboratory Medicine, Hangzhou Medical College, Hangzhou, ChinaKey Laboratory of Biomarkers and In Vitro Diagnosis Translation of Zhejiang Province, Hangzhou, ChinaDepartment of Clinical Laboratory, Laboratory Medicine Center, Zhejiang Provincial People’s Hospital, Hangzhou Medical College, Hangzhou, ChinaUltrafast, portable, and inexpensive molecular diagnostic platforms are critical for clinical diagnosis and on-site detection. There are currently no available real-time polymerase chain reaction (PCR) devices able to meet the demands of point-of-care testing, as the heating and cooling processes cannot be avoided. In this study, the dual temperature modules were first designed to process microfluidic chips automatically circulating between them. Thus, a novel ultrafast molecular diagnostic real-time PCR device (approximately 18 and 23 min for DNA and RNA detection, respectively) with two channels (FAM and Cy5) for the detection of 12 targets was developed. The device contained three core functional components, including temperature control, optics, and motion, which were integrated into a portable compact box. The temperature modules accurately control temperature in rapid thermal cycles with less than ±0.1 °C, ±1 °C and ±0.5 °C for the temperature fluctuation, uniformity, and error of indication, respectively. The average coefficient of variation (CV) of the fluorescence intensity (FI) for all 12 wells was 2.3% for FAM and 2.7% for Cy5. There was a good linear relationship between the concentrations of fluorescent dye and the FIs of FAM and Cy5(R2 = 0.9990 and 0.9937), and the average CVs of the Ct values calculated by the embedded software were 1.4% for FAM and Cy5, respectively. The 100 double-blind mocked sputum and 249 clinical stool samples were analyzed by the ultrafast real-time PCR device in comparison with the DAAN Gene SARS-CoV-2 kit run on the ABI 7500 instrument and Xpert C. difficile/Epi, respectively. Among the 249 stool samples, the ultrafast real-time PCR device detected toxigenic C. difficile in 54 samples (54/249, 21.7%) with a specificity and positive predictive values of 99.0 and 96.3%, which were higher than the Xpert C. difficile/Epi values of 94.4 and 88.1% (p > 0.05). The ultrafast real-time PCR device detected 15 SARS-CoV-2 positive samples, which has a 100% concordance with that obtained by the DAAN Gene SARS-CoV-2 kit. This study demonstrated that the ultrafast real-time PCR device integrated with microfluidic chips and dual temperature modules is an ultrafast, reliable, easy-to-use, and cost-effective molecular diagnostic platform for clinical diagnosis and on-site testing, especially in resource-limited settings.https://www.frontiersin.org/articles/10.3389/fbioe.2022.895236/fullmolecular diagnosticultrafastmicrofluidic chipdual temperature modulesperformance evaluation |
| spellingShingle | Shan Lin Shan Lin Xiaojun Song Kun Zhu Quanyu Shao Yinhang Chen Wei Cheng Zhijing Lei Yu Chen Yu Chen Yun Luo Dazhi Jin Dazhi Jin Dazhi Jin Performance Evaluation of a Novel Ultrafast Molecular Diagnostic Device Integrated With Microfluidic Chips and Dual Temperature Modules Frontiers in Bioengineering and Biotechnology molecular diagnostic ultrafast microfluidic chip dual temperature modules performance evaluation |
| title | Performance Evaluation of a Novel Ultrafast Molecular Diagnostic Device Integrated With Microfluidic Chips and Dual Temperature Modules |
| title_full | Performance Evaluation of a Novel Ultrafast Molecular Diagnostic Device Integrated With Microfluidic Chips and Dual Temperature Modules |
| title_fullStr | Performance Evaluation of a Novel Ultrafast Molecular Diagnostic Device Integrated With Microfluidic Chips and Dual Temperature Modules |
| title_full_unstemmed | Performance Evaluation of a Novel Ultrafast Molecular Diagnostic Device Integrated With Microfluidic Chips and Dual Temperature Modules |
| title_short | Performance Evaluation of a Novel Ultrafast Molecular Diagnostic Device Integrated With Microfluidic Chips and Dual Temperature Modules |
| title_sort | performance evaluation of a novel ultrafast molecular diagnostic device integrated with microfluidic chips and dual temperature modules |
| topic | molecular diagnostic ultrafast microfluidic chip dual temperature modules performance evaluation |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.895236/full |
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