Recombinase polymerase amplification combined with a lateral flow dipstick for rapid and visual detection of Plasmodium vivax
10.3969/j.issn.1002-2694.2025.00.078
- VernacularTitle:基于重组酶聚合酶等温扩增的间日疟快速可视化检测研究
- Author:
Shi-hui LI
1
;
Chun-hua GAO
1
;
Fu-rong WEI
1
;
Duo-quan WANG
1
;
Xiao-kai JIA
1
;
Jing ZHANG
1
;
Ying WANG
1
;
Feng SHI
1
Author Information
1. 中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心),国家卫生健康委员会寄生虫病原与媒介生物学重点实验室,世界卫生组织热带病合作中心,科技部国家级热带病国际研究中心,上海 200025
- Publication Type:Journal Article
- Keywords:
Plasmodium vivax;
recombinase polymerase amplification;
lateral flow dipstick;
rapid detection
- From:
Chinese Journal of Zoonoses
2025;41(4):413-418
- CountryChina
- Language:Chinese
-
Abstract:
To achieve rapid and visual detection of Plasmodium vivax,a detection method based on recombinase polymerase amplification(RPA)technology and lateral flow dipstick(LFD)was established and evaluated.Targeting the conserved sequence of the P.vivax 18S rRNA gene(GenBank:DQ660817.1)as the target sequence,primers and probes were designed with Primer Premier 5,and the P.vivax recombinant plasmid(pUCPv)was constructed as the standard.A sensitive and specific RPA-LFD-based rapid visual detection method for P.vivax nucleic acids was established.The plasmid standard was serially diluted 10-fold to concentrations of 1×103,1×102,1×101,1×10?,and 1×10?1 copies/μL for sensitivity testing.To evaluate specificity,whole blood DNA samples from patients infected with Plasmodium falciparum,Plasmodium malariae,Plasmodium ovale,or Leishmania donovani,as well as healthy participants,were tested by RPA-LFD.Additionally,The assay′s accuracy was evaluated by testing whole blood DNA samples from 24 confirmed P.vivax-infected patients.This study successfully established a sensitive,specific,and rapid visual RPA-LFD method for detecting P.vivax nucleic acids.The assay can complete P.vivax detection within 20 minutes under isothermal conditions at 39 ℃,achieving a sensitivity of 1 copy/μL.There is no significant cross reaction with parasites such as other Plasmodium species and L.donovani,and the specificity is 100%.All 24 DNA samples from confirmed P.vivax patients were detected,showing a 100%detection rate.The developed RPA-LFD assay exhibits excellent sensitivity and specificity,requires only simple heating equipment,and is user-friendly.This rapid visual detection method is particularly suitable for P.vivax screening in low-resource settings.
