Sensitive and rapid detection of Schistosoma japonicum DNA by loop-mediated isothermal amplification (LAMP)

Xu, Jin; Rong, R.; Zhang, H.Q.; Shi, Chang-Jun; Zhu, Xingquan; Xia, Chao-Ming

doi:10.1016/j.ijpara.2009.08.010

Cited by 134 publications

(117 citation statements)

References 27 publications

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“…26 Unlike PCR, LAMP does not require amplification cycles by thermocycling or amplicon detection by electrophoresis. Given these features, LAMP is potentially useful for work in the field and has already used in rural laboratories in developing areas for the diagnosis of tuberculosis.…”

Section: Discussionmentioning

confidence: 99%

“…The LAMP, initially described in 2000, 24 has rapidly gained acceptance for detection of a variety of infectious agents including Plasmodium falciparum and S. japonicum . 25,26 This technique does not require complex equipment for DNA amplification or for amplicon detection, 27 and is potentially suitable for molecular monitoring in basic laboratory facilities. 28 In the present study, we describe the development of two new LAMP assays for detecting S. haematobium and S. mansoni DNA in biological materials and their sensitivity for detecting infected snails.…”

Section: Introductionmentioning

confidence: 99%

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Detection of Schistosoma mansoni and Schistosoma haematobium DNA by Loop-Mediated Isothermal Amplification: Identification of Infected Snails from Early Prepatency

Abbasi¹,

King²,

Muchiri³

et al. 2010

The American Society of Tropical Medicine and Hygiene

105

126

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Monitoring post-control transmission of schistosomes by examining humans becomes less effective as infection rates among humans decrease. Molecular monitoring of prepatent schistosome infection in snails by the polymerase chain reaction (PCR) has been used for studying human-to-snail transmission, and snail prepatent infection rates were found to correspond to infection prevalence and average intensity in human populations contacting the sites studied. We have now developed loop-mediated isothermal amplification (LAMP) assays for identifying Schistosoma mansoni and S. haematobium to facilitate large-scale evaluation of post-intervention transmission potential. LAMP primers were designed based on the Sm1-7 and DraI repeated sequences of the corresponding schistosomes, and amplification by LAMP of these 121-basepair highly abundant sequences provided a detection sensitivity of 0.1 fg of genomic DNA. When these LAMP assays were applied for examining infected laboratory snails, it was possible to identify infection from the first day after exposure to miracidia. The potential advantages of these assays are discussed.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Detection of Schistosoma mansoni and Schistosoma haematobium DNA by Loop-Mediated Isothermal Amplification: Identification of Infected Snails from Early Prepatency

Abbasi¹,

King²,

Muchiri³

et al. 2010

The American Society of Tropical Medicine and Hygiene

105

126

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“…DNA-based detection methods provide a viable alternative to some of the commonly used tests for the diagnosis of schistosomiasis japonica Gordon et al, 2011;Xu et al, 2010). However, it is important to consider the value of testing clinical samples using different bodily fluids with DNA detection-based assays.…”

Section: Discussionmentioning

confidence: 99%

“…The assay detected target sequences in different sources of Schistosoma DNA isolated from adult worms, schistosomules and eggs; it is highly sensitive and can detect very low amounts of parasite DNA (Lier et al, 2006;Xia et al, 2009;Xu et al, 2010). The specificity of the assay is high, with no cross reactivity found when tested using DNA from other helminth parasites as template, nor was there any crossreactivity with host DNA.…”

Section: Accepted Manuscriptmentioning

confidence: 99%

Optimisation of a droplet digital PCR assay for the diagnosis of Schistosoma japonicum infection: A duplex approach with DNA binding dye chemistry

Weerakoon

Gordon

Gobert

et al. 2016

Journal of Microbiological Methods

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Schistosomiasis is a chronically debilitating helminth infection with a significant socioeconomic and public health impact. Accurate diagnostics play a pivotal role in achieving current schistosomiasis control and elimination goals. However, many of the current diagnostics procedures, which rely on detection of schistosome eggs, have major limitations including lack of accuracy and the inability to detect pre-patent infections. DNA-based detection methods provide a viable alternative to the current tests commonly used for schistosomiasis diagnosis. Here we describe the optimisation of a novel droplet digital PCR (ddPCR) duplex assay for the diagnosis of Schistosoma japonicum which provides improved detection sensitivity and specificity. The assay involves the amplification of two specific and abundant target gene sequences in S. japonicum; a retrotransposon (SjR2) and a portion of a mitochondrial gene (nad1). The assay detected target sequences in different sources of schistosome DNA isolated from adult worms, schistosomules and eggs, and exhibits a high level of specificity, thereby representing an ideal tool for the detection of low levels of parasite DNA in different clinical samples including parasite cell free DNA in the host circulation and other bodily fluids. Moreover, being quantitative, the assay can be used to determine parasite infection intensity and, could provide an important tool for the detection of low intensity infections in low prevalence schistosomiasis-endemic areas.

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“…The low tech PCR approach, loop mediated isothermal amplification (LAMP) developed by Notomi et al 96 was recently used for the detection of S. japonicum DNA in rabbit fecal and serum samples, as well as in 50 human serum samples 97 . The analytical test evaluation revealed high analytical sensitivity, detecting 0.08fg of DNA, and no cross-reaction was found with S. mansoni and Clonorchis sinensis.…”

mentioning

confidence: 99%

Diagnosing schistosomiasis: where are we?

Gomes

Enk

Rabello

2014

Rev. Soc. Bras. Med. Trop.

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In light of the World Health Organization's initiative to extend schistosomiasis morbidity and mortality control programs by including a disease elimination strategy in low endemic settings, this paper reviews diagnostic tools described during the last decades and provide an overview of ongoing efforts in making an efficient diagnostic tool available worldwide. A literature search on PubMed using the search criteria schistosomiasis and diagnosis within the period from 1978 to 2013 was carried out. Articles with abstract in English and that used laboratory techniques specifically developed for the detection of schistosomiasis in humans were included. Publications were categorized according to the methodology applied (parasitological, immunological, or molecular) and stage of development (in house development, limited field, or large scale field testing). The initial research generated 4,535 publications, of which only 643 met the inclusion criteria. The vast majority (537) of the publications focused on immunological techniques; 81 focused on parasitological diagnosis, and 25 focused on molecular diagnostic methods. Regarding the stage of development, 307 papers referred to in-house development, 202 referred to limited field tests, and 134 referred to large scale field testing. The data obtained show that promising new diagnostic tools, especially for Schistosoma antigen and deoxyribonucleic acid (DNA) detection, which are characterized by high sensitivity and specificity, are being developed. In combination with international funding initiatives these tools may result in a significant step forward in successful disease elimination and surveillance, which is to make efficient tests accessible and its large use self-sustainable for control programs in endemic countries.

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Sensitive and rapid detection of Schistosoma japonicum DNA by loop-mediated isothermal amplification (LAMP)

Cited by 134 publications

References 27 publications

Detection of Schistosoma mansoni and Schistosoma haematobium DNA by Loop-Mediated Isothermal Amplification: Identification of Infected Snails from Early Prepatency

Detection of Schistosoma mansoni and Schistosoma haematobium DNA by Loop-Mediated Isothermal Amplification: Identification of Infected Snails from Early Prepatency

Optimisation of a droplet digital PCR assay for the diagnosis of Schistosoma japonicum infection: A duplex approach with DNA binding dye chemistry

Diagnosing schistosomiasis: where are we?

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