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

<p>Abstract</p> <p>Background</p> <p>A 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 poly...

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Main Authors: Lee Pei-Wen, Ji Dar-Der, Liu Chia-Tai, Rampao Herodes S, do Rosario Virgilio E, Lin I-Feng, Shaio Men-Fang
Format: Article
Language:English
Published: BMC 2012-12-01
Series:Malaria Journal
Subjects:
Online Access:http://www.malariajournal.com/content/11/1/408
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author Lee Pei-Wen
Ji Dar-Der
Liu Chia-Tai
Rampao Herodes S
do Rosario Virgilio E
Lin I-Feng
Shaio Men-Fang
author_facet Lee Pei-Wen
Ji Dar-Der
Liu Chia-Tai
Rampao Herodes S
do Rosario Virgilio E
Lin I-Feng
Shaio Men-Fang
author_sort Lee Pei-Wen
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>A 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.</p> <p>Method</p> <p>During 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.</p> <p>Results</p> <p>On 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.</p> <p>Conclusions</p> <p>HRP-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.</p>
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spelling doaj.art-af9b4efc015543509ada5bad8122d8ce2022-12-22T02:41:10ZengBMCMalaria Journal1475-28752012-12-0111140810.1186/1475-2875-11-408Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São ToméLee Pei-WenJi Dar-DerLiu Chia-TaiRampao Herodes Sdo Rosario Virgilio ELin I-FengShaio Men-Fang<p>Abstract</p> <p>Background</p> <p>A 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.</p> <p>Method</p> <p>During 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.</p> <p>Results</p> <p>On 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.</p> <p>Conclusions</p> <p>HRP-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.</p>http://www.malariajournal.com/content/11/1/408MalariaMicroscopyRapid diagnostic testNested polymerase chain reactionLoop-mediated isothermal amplificationTreatment follow-up
spellingShingle Lee Pei-Wen
Ji Dar-Der
Liu Chia-Tai
Rampao Herodes S
do Rosario Virgilio E
Lin I-Feng
Shaio Men-Fang
Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé
Malaria Journal
Malaria
Microscopy
Rapid diagnostic test
Nested polymerase chain reaction
Loop-mediated isothermal amplification
Treatment follow-up
title Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé
title_full Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé
title_fullStr Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé
title_full_unstemmed Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé
title_short Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé
title_sort application of loop mediated isothermal amplification for malaria diagnosis during a follow up study in sao tome
topic Malaria
Microscopy
Rapid diagnostic test
Nested polymerase chain reaction
Loop-mediated isothermal amplification
Treatment follow-up
url http://www.malariajournal.com/content/11/1/408
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