Development of Loop-Mediated Isothermal Amplification Assay for Detection of
            <i>Entamoeba histolytica</i>

Liang, Shih Yu; Chan, Yun Hsien; Hsia, Kan Tai; Lee, Jing Lun; Kuo, Ming Chu; Hwa, Kuo-Yuan; Chan, Chi Wen; Chiang, Ting Yi; Chen, Jung‐Sheng; Wu, Fengkai; Ji, Dar Der

doi:10.1128/jcm.00105-09

Cited by 62 publications

(42 citation statements)

References 23 publications

(25 reference statements)

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“…The positive LAMP reaction was detected by observing the fluorescence in the reaction tube via naked eye under a portable UV lamp [22]. In this study, an interval of 10 min was set to determine the threshold time of the LAMP reaction.…”

Section: Methodsmentioning

confidence: 99%

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

Lee¹,

Liu³

et al. 2012

Malar J

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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.

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Section: Methodsmentioning

confidence: 99%

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

Lee¹,

Liu³

et al. 2012

Malar J

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“…A change from polymerase chain reaction to loop-mediated isothermal amplication (LAMP) has certain advantages because temperature cycling is not necessary. A number of other amplification systems which do not require temperature cycling have also been developed (Compton, 1991;Guatelli et al, 1990;Walker et al, 1992a,b;Abramowitz, 1996;Vrana, 1996) and applied to parasite detection (Matovu et al, 2010a), but LAMP is currently the most widely used alternative to PCR (Kuboki et al, 2003;Thekisoe et al, 2005;Alhassan et al, 2007;Han et al, 2007;Karanis et al, 2007;Guan et al, 2008;Njiru et al, 2008a,b;Peyrefitte et al, 2008;Aonuma et al, 2009;Liang et al, 2009;Nkouawa et al, 2009;Takagi et al, 2009;Abbasi et al, 2010;Adams et al, 2010;Iseki et al, 2010;Kumagai et al, 2010;Lau et al, 2010;Matovu et al, 2010b;Ndung'u et al, 2010;Polley et al, 2010;Poschl et al, 2010;Thekisoe et al, 2010;Wang et al, 2010;Njiru, 2011). LAMP and these other alternatives to PCR are still based on enzymatic reactions however, so the transport and storage of reagents at below room temperature is a requirement and a challenge for field application.…”

Section: Problems With Pcr; Lamp and Other Potential Solutionsmentioning

confidence: 98%

Molecular diagnosis of infections and resistance in veterinary and human parasites

Hunt

2011

Veterinary Parasitology

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“…The white turbidity can be observed by the naked eye or measured with a turbidimeter. LAMP has been used to detect protozoan parasites such as Entamoeba, Babesia, Leishmania, Trypanosoma, Theileria, Giardia, and Cryptosporidium in humans and animals (Alhassan et al 2007;Bakheit et al 2008;Guan et al 2008;Liu et al 2008;Njiru et al 2008;Liang et al 2009;Plutzer and Karanis 2009;Laohasinnarong et al 2011). LAMP has also been successfully developed for species-specific detection of human malaria parasites (Poon et al 2006;Han et al 2007;Chen et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Molecular test for vivax malaria with loop-mediated isothermal amplification method in central China

Gao

Zhou

et al. 2011

Parasitol Res

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Vivax malaria has the highest incidence in central China. High-throughput and cost-effective testing methods are essential for vivax malaria diagnosis in this area. Loop-mediated isothermal amplification (LAMP) is an established nucleic acid amplification method, which provides a promising platform for the molecular detection of malaria parasites in development countries. This study was performed to compare the LAMP method, the nested PCR-based method, and microscopic examinations in diagnosing vivax malaria. LAMP detected vivax malaria parasites in 160 of 164 microscopically positive samples (sensitivity, 97.6%), whereas nested PCR detected Plasmodium vivax in 162 of 164 samples (sensitivity, 98.8%). No false-positive results were obtained by LAMP or nested PCR among fever-positive and healthy samples. The sensitivities and specificities of the two molecular tests were consistently high. In addition, the reproducibility of the LAMP assays using water bath, which reduced the cost substantially. LAMP method is an accurate, rapid, simple, and cost-effective method that may be useful for diagnosis in field diagnoses instead of nested PCR. This method is feasible to diagnose vivax malaria parasite in endemic areas of central China.

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Development of Loop-Mediated Isothermal Amplification Assay for Detection of Entamoeba histolytica

Cited by 62 publications

References 23 publications

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

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

Molecular diagnosis of infections and resistance in veterinary and human parasites

Molecular test for vivax malaria with loop-mediated isothermal amplification method in central China

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