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Volume 39 Issue 1
Jan.  2023
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LIU Jian-li, GAO Jing, JIANG Peng-chong, . Establishment of an automatic detection method for Middle East respira-tory syndrome coronavirus with an integrated microfluidic chip[J]. Chinese Journal of Public Health, 2023, 39(1): 92-97. doi: 10.11847/zgggws1139190
Citation: LIU Jian-li, GAO Jing, JIANG Peng-chong, . Establishment of an automatic detection method for Middle East respira-tory syndrome coronavirus with an integrated microfluidic chip[J]. Chinese Journal of Public Health, 2023, 39(1): 92-97. doi: 10.11847/zgggws1139190

Establishment of an automatic detection method for Middle East respira-tory syndrome coronavirus with an integrated microfluidic chip

doi: 10.11847/zgggws1139190
  • Received Date: 2020-05-17
    Available Online: 2023-01-09
  • Publish Date: 2023-01-31
  •   Objective   To develop an automatic detection method for Middle East respiratory syndrome coronavirus (MERS-CoV) based on integrated microfluidic chip platform.   Methods  The pseudovirus particle of MERS-CoV was prepared as positive reference and primers and probe were designed for the ORF1b gene region of the virus. Integrated microfluidic chips were created in accordance with the whole procedure of real-time reverse transcription polymerase chain reaction (RT-PCR), including viral lysis, nucleic acid extraction, amplification and real-time fluorescence detection. The performance of the chip was monitored by the internal reference gene embedded on the chip. The sensitivity, specificity and repeatability of the chip were evaluated using the MERS-CoV pseudovirus.   Results  The internal reference gene on the microfluidic chip could be amplified and detected stably. The automated detection was sensitive enough to detect as few as 100 copies of MERS-CoV in the original sample, which was more sensitive than normal real-time RT-PCR with the detection limitation of 500 copies. A cycle threshold (Ct) value of 39.80 ± 0.53, with a coefficient variation of 1.34%, was obtained when the test was repeated six times with 100 copies of pseudovirus. No signal was observed in automated tests for other coronaviruses and human genes.   Conclusion  The established automatic detection method with microfluidic chip is of good stability, sensitivity, and specificity and the method is fast and easy to operate, and is not limited by experimental sites. The method could be used for rapid detection of MERS-CoV in field investigation.
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  • [1]
    董家潇, 李修政, 张忠伟, 等. SARS - COV和MERS - COV以及SARS-COV-2的研究进展[J]. 中国病毒病杂志, 2021, 11(5): 381 – 387. doi: 10.16505/j.2095-0136.2021.0050
    [2]
    World Health Organization. MERS situation update, March 2022[EB/OL]. (2022 – 02 – 20). http://www.emro.who.int/health-topics/mers-cov/mers-outbreaks.html.
    [3]
    陆柔剑, 武婕, 牛培华, 等. 中国首例输入性中东呼吸综合征病例实验室确诊方法比较[J]. 中华实验和临床病毒学杂志, 2015, 29(3): 193 – 195. doi: 10.3760/cma.j.issn.1003-9279.2015.03.001
    [4]
    于海江, 于洋洋, 于栋, 等. SARS-CoV-2、SARS-CoV-1及MERS-CoV的特征比较分析[J]. 基因组学与应用生物学, 2020, 39(9): 4400 – 4404. doi: 10.13417/j.gab.039.004400
    [5]
    王晓欢, 严延生, 张智芳, 等. 中东呼吸综合征[J]. 中国人兽共患病学报, 2020, 36(6): 496 – 502.
    [6]
    World Health Organization. Laboratory testing for middle east respiratory syndrome coronavirus-revised, interim guidance[EB/OL]. (2018 – 01 – 15). https://www.who.int/publications/i/item/10665-259952.
    [7]
    Taki K, Yokota I, Fukumoto T, et al. SARS - CoV - 2 detection by fluorescence loop-mediated isothermal amplification with and without RNA extraction[J]. Journal of Infection and Chemotherapy, 2021, 27(2): 410 – 412. doi: 10.1016/j.jiac.2020.10.029
    [8]
    Wei S, Kohl E, Djandji A, et al. Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction[J]. Scientific Reports, 2021, 11(1): 2402.
    [9]
    Pearson JD, Trcka D, Lu SY, et al. Comparison of SARS - CoV - 2 indirect and direct RT - qPCR detection methods[J]. Virology Journal, 2021, 18(1): 99. doi: 10.1186/s12985-021-01574-4
    [10]
    Beltrán-Pavez C, Alonso-Palomares LA, Valiente-Echeverría F, et al. Accuracy of a RT - qPCR SARS - CoV - 2 detection assay without prior RNA extraction[J]. Journal of Virological Methods, 2021, 287: 113969. doi: 10.1016/j.jviromet.2020.113969
    [11]
    Shabani E, Dowlatshahi S, Abdekhodaie MJ. Laboratory detec-tion methods for the human coronaviruses[J]. European Journal of Clinical Microbiology and Infectious Diseases, 2021, 40(2): 225 – 246. doi: 10.1007/s10096-020-04001-8
    [12]
    Huang P, Wang HL, Cao ZG, et al. A rapid and specific assay for the detection of MERS - CoV[J]. Frontiers in Microbiology, 2018, 9: 1101. doi: 10.3389/fmicb.2018.01101
    [13]
    Shahrajabian MH, Sun WL, Cheng Q. Different methods for molecular and rapid detection of human novel coronavirus[J]. Current Pharmaceutical Design, 2021, 27(25): 2893 – 2903. doi: 10.2174/1381612827666210604114411
    [14]
    Ye X, Li Y, Wang LJ, et al. All-in-one microfluidic nucleic acid diagnosis system for multiplex detection of sexually transmitted pathogens directly from genitourinary secretions[J]. Talanta, 2021, 221: 121462. doi: 10.1016/j.talanta.2020.121462
    [15]
    Liu Q, Zhang XL, Chen LP, et al. A sample - to - answer labdisc platform integrated novel membrane - resistance valves for detection of highly pathogenic avian influenza viruses[J]. Sensors and Actuators B: Chemical, 2018, 270: 371 – 381. doi: 10.1016/j.snb.2018.05.044
    [16]
    Chen JG, Xu YC, Yan H, et al. Sensitive and rapid detection of pathogenic bacteria from urine samples using multiplex recom-binase polymerase amplification[J]. Lab on a Chip, 2018, 18(16): 2441 – 2452. doi: 10.1039/C8LC00399H
    [17]
    Kim MN, Ko YJ, Seong MW, et al. Analytical and clinical validation of six commercial middle East respiratory syndrome coronavirus RNA detection kits based on real - time reverse - transcription PCR[J]. Annals of Laboratory Medicine, 2016, 36(5): 450 – 456. doi: 10.3343/alm.2016.36.5.450
    [18]
    迟航, 郑学星, 盖微微, 等. 中东呼吸综合征冠状病毒实时荧光定量PCR检测方法的建立[J]. 中国病原生物学杂志, 2014, 9(8): 673 – 677. doi: 10.13350/j.cjpb.140801
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    • Receive:  2020-05-17
    • Online:  2023-01-09
    • Published:  2023-01-31

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