Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé

Malaria Journal

Research

Herodes S Rampao¹  I-Feng Lin²  Virgilio E do Rosario³  Dar-Der Ji⁴  Chia-Tai Liu⁵  Pei-Wen Lee⁵  Men-Fang Shaio⁶ 

[1] Centro National de Endemias, São Tomé, Taipei, Democratic Republic of São Tomé and Príncipe, Taiwan;Institute of Public Health, National Yang-Ming University, Taipei, Taiwan;Instituto de Higiene e Medicina Tropical/Universidade Nova de Lisboa, Lisbon, Portugal;Research and Diagnostic Center, Centers for Disease Control, Taipei, Taiwan;Department of Tropical Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan;The Anti-Malaria Team of Taiwan in São Tomé and Príncipe, Taipei, São Tomé and Princípe, Taiwan;The Anti-Malaria Team of Taiwan in São Tomé and Príncipe, Taipei, São Tomé and Princípe, Taiwan;Department of Tropical Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan;

关键词: Malaria;    Microscopy;    Rapid diagnostic test;    Nested polymerase chain reaction;    Loop-mediated isothermal amplification;    Treatment follow-up;   

DOI  :  10.1186/1475-2875-11-408

 received in 2012-09-21, accepted in 2012-12-03,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundA reliable and simple test for the detection of malaria parasite is crucial in providing effective treatment and therapeutic follow-up, especially in malaria elimination programmes. A comparison of four methods, including nested polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) were used for the malaria diagnosis and treatment follow-up in São Tomé and Príncipe, during a successful pre-elimination campaign.MethodDuring the period September to November 2009, blood samples from 128 children (five to 14 years old) with temperature ≥38°C (tympanic) in the District of Agua Grande were examined using four different methods, i.e., histidine-rich protein 2 (HRP-2) based rapid diagnostic tests (HRP-2-RDTs), optical microscopy, nested PCR, and LAMP. First-line treatment with artesunate-amodiaquine was given for uncomplicated malaria and intravenous quinine was given for complicated malaria. Children with persistent positivity for malaria by microscopy, or either by nested PCR, or by LAMP on day 7 were given second-line treatment with artemether-lumefantrine. Treatment follow-up was made weekly, for up to four weeks.ResultsOn day 0, positive results for HRP-2-RDTs, microscopy, nested PCR, and LAMP, were 68(53%), 47(37%), 64(50%), and 65(51%), respectively. When nested PCR was used as a reference standard, only LAMP was comparable; both HRP-2-RDTs and microscopy had moderate sensitivity; HRP-2-RDTs had poor positive predictive value (PPV) and a moderate negative predictive value (NPV) for the treatment follow-up. Seventy-one children with uncomplicated malaria and eight children with complicated falciparum malaria were diagnosed based on at least one positive result from the four tests as well as clinical criteria. Twelve of the 79 children receiving first-line treatment had positive results by nested PCR on day 7 (nested PCR-corrected day 7 cure rate was 85%). After the second-line treatment, nested PCR/LAMP-corrected day 28 cure rate was 83% for these 12 children.ConclusionsHRP-2-RDTs have similar sensitivity as microscopy but less specificity. However, as compared to nested PCR, the poor sensitivity of HRP-2-RDTs indicates that low parasitaemia may not be detected after treatment, as well as the low specificity of HRP-2-RDTs indicates it cannot be applied for treatment follow-up. LAMP has similar sensitivity and specificity to nested PCR. With high PPV and NPV, LAMP is simpler and faster as compared to nested PCR with the advantage of detecting low parasitaemia becoming a potential point-of-care test for treatment follow-up.

【 授权许可】

Unknown   
© Lee et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.