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A Rapid On-Site Detection Method for Plasmodium falciparum Carried by Mosquitoes Using Disc Microfluidic Isothermal Amplification
Ning Jiang
Ning Jiang
Department of Pathogenic Biology, Basic Medical College, Naval Medical University, Shanghai, China;
Clinical Laboratory, Naval Medical Centre, Naval Medical University, Shanghai, China;
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Clinical Laboratory, Naval Medical Centre, Naval Medical University, Shanghai, China;
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Jie Bai
Jie Bai
Department of Pathogenic Biology, Basic Medical College, Naval Medical University, Shanghai, China;
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Wenqi Shan
Wenqi Shan
Department of Naval Medicine, Naval Medical University, Shanghai, China;
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Qiuming Zhou
Qiuming Zhou
Department of Naval Medicine, Naval Medical University, Shanghai, China;
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Haowei Dong
Haowei Dong
Department of Pathogenic Biology, Basic Medical College, Naval Medical University, Shanghai, China;
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Hao Yuan
Hao Yuan
Department of Naval Medicine, Naval Medical University, Shanghai, China;
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Wenbing Zhong
Wenbing Zhong
Haikou Center for Disease Control and Prevention, Hainan, China;
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Xiangyu Li
Xiangyu Li
Department of Pathogenic Biology, Basic Medical College, Naval Medical University, Shanghai, China;
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Yajun Ma
Yajun Ma
Department of Naval Medicine, Naval Medical University, Shanghai, China;
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Ping Li
15109827722@163.com
Ping Li
Haikou Center for Disease Control and Prevention, Hainan, China;
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Heng Peng
pengheng0923@126.com
Heng Peng
Department of Pathogenic Biology, Basic Medical College, Naval Medical University, Shanghai, China;
Key Laboratory of Biosafety Defense, Naval Medical University, Ministry of Education, China
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Key Laboratory of Biosafety Defense, Naval Medical University, Ministry of Education, China
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Rapidly identifying Anopheles-carrying malaria parasites is crucial for imported malaria prevention. However, suitable methods still lack quick detection in limited-resource situations. In this study, disc microfluidic isothermal amplification integrating loop-mediated isothermal amplification (LAMP) and microfluidic chip technology were applied to develop rapid and precise detection with low resource requirements. Primer set EMP1G2, which is specific to Plasmodium falciparum (P. falciparum) erythrocyte membrane protein 1, and primer set 18sG2, which is specific to ribosomal 18s subunit RNA, were screened for optimal LAMP-specific primer sets. The minimum detection limits were 125 copies/µL for the EMP1G2 and 6,562 copies/µL for the 18sG2. Subsequently, optimal primer sets were evaluated for specificity with nucleic acid from other mosquito-borne pathogens and arthropod vectors. No nonspecific amplification was observed in optimal amplification-specific primer sets with the DNA of Anopheles mosquitoes and morphologically similar arthropods or with the copy DNA of Zika virus, yellow fever virus, or dengue virus 1. The detection method was evaluated in a simulated scenario and demonstrated a robust capacity for rapid on-site detection. Additionally, polymerase chain reaction (PCR) and quantitative real-time PCR methods were compared using this method. In this study, a rapid detection method based on disc microfluidic isothermal amplification was developed that could be used to detect P. falciparum carried by mosquitoes in a field setting under limited resource conditions.
Author Notes
Financial support: This work was supported by the sixth round of the
Disclosures: The nucleic acids of pathogen and mosquito samples involved in this study do not raise any ethical concerns; therefore, ethics approval for this study was not required.
Authors’ contributions: Conceptualization, P. Li and H. Peng; methodology, N. Jiang and J. Bai; validation, W. Shan; formal analysis, N. Jiang, Q. Zhou, and H. Dong; investigation, X. Li; resources, Y. Ma and H. Peng; data curation, N. Jiang and H. Yuan; writing—original draft preparation, N. Jiang; writing—review and editing, P. Li and H. Peng; visualization, N. Jiang and W. Zhong; supervision, Y. Ma and W. Zhong; project administration, H. Peng and Y. Ma; funding acquisition, H. Peng and Y. Ma. All authors have read and agreed to the published version of the manuscript.
Data availability: The data are available from the corresponding author on reasonable request.
Current contact information: Ning Jiang, Clinical Laboratory, Naval Medical Centre, Naval Medical University, Shanghai, China, E-mail: jiangning990921@163.com. Jie Bai, Haowei Dong, Xiangyu Li, and Heng Peng, Department of Pathogenic Biology, Basic Medical College, Naval Medical University, Shanghai, China, E-mails: 13761503780@163.com, wsnd19951031@163.com, jack_lee@smmu.edu.cn, and pengheng0923@126.com. Wenqi Shan, Qiuming Zhou, Hao Yuan, and Yajun Ma, Department of Naval Medicine, Naval Medical University, Shanghai, China, E-mails: wenqi_shan@163.com, zhouqm0201@163.com, yhao07@126.com, and yajun_ma@163.com. Wenbing Zhong and Ping Li, Haikou Center for Disease Control and Prevention, Hainan, China, E-mails: zhongwenbing66@163.com and 15109827722@163.com.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 461 | 461 | 461 |
| Full Text Views | 37 | 37 | 37 |
| PDF Downloads | 26 | 26 | 26 |