Comparison of nucleic acid sequence-based amplification and loop-mediated isothermal amplification for diagnosis of human African trypanosomiasis

Mugasa, Claire M.; Katiti, Diana; Boobo, Alex; Lubega, George W.; Schallig, Henk D. F. H.; Matovu, Enock

doi:10.1016/j.diagmicrobio.2013.06.025

Cited by 17 publications

(9 citation statements)

References 29 publications

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“…This assay was further simplified by using membrane oligochromatography (OC) for rapid detection of RNA following amplification (Mugasa et al, 2009). A recent comparison of a LAMP assay and the NASBA-OC assay showed the latter to have a higher level of positive and negative agreement with microscopy for detection of T. b. gambiense (Mugasa et al, 2014). The NASBA-OC assay used in this study was also compared to direct microscopy for detection of T. brucei in clinical specimens and showed a significantly higher sensitivity (73% vs. 57% for microscopy) compared to a composite gold standard (Mugasa et al, 2009).…”

Section: Diagnostic Modalitiesmentioning

confidence: 99%

Molecular Diagnostics in the Diagnosis of Parasitic Infection

Pritt¹

2015

Methods in Microbiology

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Section: Diagnostic Modalitiesmentioning

confidence: 99%

Molecular Diagnostics in the Diagnosis of Parasitic Infection

Pritt¹

2015

Methods in Microbiology

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“…However, this method is time consuming, dependent on operator expertise, unreliable for mixed infections, fails to detect immature infections and, in the case of African trypanosomes, is only useful for distinguishing between parasites to the level of subgenus (Ouma et al 2000;Gibson, 2009;Enyaru et al 2010;Auty et al 2012b;Mugasa et al 2014).…”

Section: Timeline Of Trypanosome Detectionmentioning

confidence: 99%

“…The use of isothermal amplification molecular methods, such as loop-mediated isothermal amplification and nucleic acid sequencebased amplification are becoming increasingly popular for the detection of trypanosomes as they offer simple, rapid and cheap alternatives to traditional PCR-based methods (Mugasa et al 2014;Besuschio et al 2017;Rivero et al 2017). Isothermal tests involve a single reaction in a single tube incubated at a constant temperature; therefore, these techniques do not require the expensive thermocycling equipment that is necessary for PCR Wastling and Welburn, 2011).…”

Section: An Alternative Future For Diagnostics? Isothermal Techniquesmentioning

confidence: 99%

“…In addition, the ability of these tests to amplify extremely small amounts of ParasitologyDNA mean that they are highly prone to contamination. Developing simplified 'kit' forms of these techniques, and refining those already available, may yield promising alternatives to sequence-based barcoding for clinical purposes (Mugasa et al 2014).…”

Section: An Alternative Future For Diagnostics? Isothermal Techniquesmentioning

confidence: 99%

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Barcoding in trypanosomes

Hutchinson

Stevens

2017

Parasitology

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Trypanosomes (genus Trypanosoma) are parasites of humans, and wild and domestic mammals, in which they cause several economically and socially important diseases, including sleeping sickness in Africa and Chagas disease in the Americas. Despite the development of numerous molecular diagnostics and increasing awareness of the importance of these neglected parasites, there is currently no universal genetic barcoding marker available for trypanosomes. In this review we provide an overview of the methods used for trypanosome detection and identification, discuss the potential application of different barcoding techniques and examine the requirements of the 'ideal' trypanosome genetic barcode. In addition, we explore potential alternative genetic markers for barcoding Trypanosoma species, including an analysis of phylogenetically informative nucleotide changes along the length of the 18S rRNA gene.

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“…Another isothermal amplification technique, known as NASBA is a method, which amplifies RNA templates, mimicking the retroviral replication mechanism, and is particularly suited for the diagnosis of RNA viruses and in detecting viable targets (Craw and Balachandran, 2012). Although NASBA has been successfully applied to the diagnosis of parasitic infections such as leishmaniasis (de Ruiter et al 2014) and trypanosomiasis (Mugasa et al 2014), it has yet to be applied to human helminths. More recently a further isothermal method, the RPA (Piepenburg et al 2006) has come to the attention of the scientific community.…”

mentioning

confidence: 99%

Focusing nucleic acid-based molecular diagnostics and xenomonitoring approaches for human helminthiases amenable to preventive chemotherapy

et al. 2016

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The current mainstay for control of the four major helminth diseases in humans (lymphatic filariasis, onchocerciasis, soiltransmitted helminthiases and schistosomiasis) is with preventive chemotherapy by mass administration of key anthelminthics. Following the London Declaration on Neglected Tropical Diseases in 2012, a roadmap for the elimination and control of these helminthiases by 2020 has been devised. With expected declines in prevalence and intensity of these infections, there is urgent need for implementing more sensitive, high-throughput and cost-effective diagnostic tools. Currently available diagnostic approaches for surveying, monitoring and evaluating helminth control programmes are based on microscopical observation of eggs/larvae, and/or detection of antibodies or parasite antigens in stool, urine or blood; all relatively low-throughput and of limited sensitivity and specificity. Newly proposed approaches for helminthiases diagnosis include the nucleic acid-based methods of (multiplex) real-time polymerase chain reaction assays, loop-mediated isothermal amplification and recombinase polymerase amplification. However, as well as sensitivity/specificity evaluation, their comparison to current 'gold standard' diagnostics and future application in individual-/community-based diagnosis, or in xenomonitoring requires consideration of relative costs, agreement of standard methods and strategic interpretation of resulting data before control/elimination programmes might best utilize molecular diagnostics to inform decision making. We review current nucleic-acid-based molecular diagnostic methods and highlight the needs and future research required to refine these tools for monitoring and evaluation of control and elimination programmes for four major human helminthiases.

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Comparison of nucleic acid sequence-based amplification and loop-mediated isothermal amplification for diagnosis of human African trypanosomiasis

Cited by 17 publications

References 29 publications

Molecular Diagnostics in the Diagnosis of Parasitic Infection

Molecular Diagnostics in the Diagnosis of Parasitic Infection

Barcoding in trypanosomes

Focusing nucleic acid-based molecular diagnostics and xenomonitoring approaches for human helminthiases amenable to preventive chemotherapy

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