Loop-mediated isothermal amplification (LAMP) assays targeting 18S ribosomal RNA genes for identifying P. vivax and P. ovale species and mitochondrial DNA for detecting the genus Plasmodium

Parasites & Vectors

Benjamin M. Rosenthal¹  Yanmei Zhang²  Xun Wang²  Hui Zhao²  Qi Yang²  Weilin Zeng²  Mengxi Duan²  Xinxin Li²  Xiaosong Li²  Zheng Xiang²  Wei Zhao²  Zhaoqing Yang²  Yaming Huang³  Jiaqi Zhang⁴  Dominique Mazier⁵  Xi Chen⁶  Yucheng Qin⁷  Maohua Pan⁷  Pien Qin⁷ 

[1] Animal Parasitic Disease Laboratory, USDA-Agricultural Research Service, 10300 Baltimore Avenue, 20705, Beltsville, MD, USA;Department of Pathogen Biology and Immunology, Kunming Medical University, 650500, Kunming, Yunnan, People’s Republic of China;Department of Pathogen Biology and Immunology, Kunming Medical University, 650500, Kunming, Yunnan, People’s Republic of China;Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, 530021, Nanning, Guangxi, People’s Republic of China;Department of Pathogen Biology and Immunology, Kunming Medical University, 650500, Kunming, Yunnan, People’s Republic of China;Zhejiang Provincial Center for Disease Control and Prevention, No. 3399 BinSheng Road, Binjiang District, 310051, Hangzhou, Zhejiang, People’s Republic of China;INSERM, CNRS, Centre d’Immunologie et des Maladies Infectieuses (CIMI), Sorbonne Université, 75013, Paris, France;Laboratory of Pathogen Biology and Immunology, Kunming Medical University, 650500, Kunming, Yunnan, People’s Republic of China;Department of Pathogen Biology and Immunology, Kunming Medical University, 650500, Kunming, Yunnan, People’s Republic of China;Shanglin County People’s Hospital, 530500, Shanglin, Guangxi, People’s Republic of China;

关键词: LAMP;    Plasmodium;    Plasmodium vivax;    Plasmodium ovale;   

DOI  :  10.1186/s13071-021-04764-9

来源: Springer

【 摘 要 】

BackgroundLoop-mediated isothermal amplification (LAMP) has been widely used to diagnose various infectious diseases. Malaria is a globally distributed infectious disease attributed to parasites in the genus Plasmodium. It is known that persons infected with Plasmodium vivax and P. ovale are prone to clinical relapse of symptomatic blood-stage infections. LAMP has not previously been specifically evaluated for its diagnostic performance in detecting P. ovale in an epidemiological study, and no commercial LAMP or rapid diagnostic test (RDT) kits are available for specifically diagnosing infections with P. ovale.MethodsAn assay was designed to target a portion of mitochondrial DNA (mtDNA) among Plasmodium spp., the five human Plasmodium species and two other assays were designed to target the nuclear 18S ribosomal DNA gene (18S rDNA) of either P. vivax or P. ovale for differentiating the two species. The sensitivity of the assays was compared to that of nested PCR using defined concentrations of plasmids containing the target sequences and using limiting dilutions prepared from clinical isolates derived from Chinese workers who had become infected in Africa or near the Chinese border with Myanmar.ResultsThe results showed that 102 copies of the mitochondrial target or 102 and 103 copies of 18S rDNA could be detected from Plasmodium spp., P. vivax and P. ovale, respectively. In 279 clinical samples, the malaria Pan mtDNA LAMP test performed well when compared with a nested PCR assay (95% confidence interval [CI] sensitivity 98.48–100%; specificity 90.75–100%). When diagnosing clinical cases of infection with P. vivax, the 18S rDNA assay demonstrated an even great sensitivity (95.85–100%) and specificity (98.1–100%). The same was true for clinical infections with P. ovale (sensitivity 90.76–99.96%; specificity 98.34–100%). Using plasmid-positive controls, the limits of detection of Malaria Pan, 18S rDNA P. vivax and 18S rDNA P. ovale LAMP were 100-, 100- and tenfold lower than those of PCR, respectively.ConclusionThe novel LAMP assays can greatly aid the rapid, reliable and highly sensitive diagnosis of infections of Plasmodium spp. transmitted among people, including P. vivax and P. ovale, cases of which are most prone to clinical relapse.Graphic abstract

【 授权许可】

CC BY