Loop-mediated isothermal amplification methods for diagnosis of visceral leishmaniasis (<i>kala-azar</i>) – a systematic review

Bezerra, Gilberto Silva Nunes; Júnior, Walter Lins Barbosa; Vieira, Amanda Virginia Batista; Xavier, Amanda Tavares; Lima-Junior, Manoel Sebastião da Costa; Xavier, Edeneide Maria; Silva, Elis Dionísio da; Leal, Nilma Cintra; Medeiros, Zulma

doi:10.1080/14737159.2020.1736564

Cited by 9 publications

(5 citation statements)

References 35 publications

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“…Alternatively, a specific PCR product targeting single or multiple species is also available, and, in general, they are able to identify subgenera and/or a group of species [ 52 ]. More recently, loop-mediated isothermal amplification (LAMP) has also been validated as an alternative method for diagnosis ( Table 1 ) [ 66 , 109 ]. Finally, for all these methods, standardization and optimization are required for use in hospitals and reference centers.…”

Section: Molecular Methods For Diagnosis Of Leishmaniasismentioning

confidence: 99%

Laboratory Diagnosis of Cutaneous and Visceral Leishmaniasis: Current and Future Methods

et al. 2020

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Leishmaniasis is a neglected tropical disease with two main clinical forms: cutaneous and visceral leishmaniasis. Diagnosis of leishmaniasis is still a challenge, concerning the detection and correct identification of the species of the parasite, mainly in endemic areas where the absence of appropriate resources is still a problem. Most accessible methods for diagnosis, particularly in these areas, do not include the identification of each one of more than 20 species responsible for the disease. Here, we summarize the main methods used for the detection and identification of leishmaniasis that can be performed by demonstration of the parasite in biological samples from the patient through microscopic examination, by in vitro culture or animal inoculation; by molecular methods through the detection of parasite DNA; or by immunological methods through the detection of parasite antigens that may be present in urine or through the detection of specific antibodies against the parasite. Potential new methods that can be applied for laboratory diagnosis of leishmaniasis are also discussed.

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Section: Molecular Methods For Diagnosis Of Leishmaniasismentioning

confidence: 99%

Laboratory Diagnosis of Cutaneous and Visceral Leishmaniasis: Current and Future Methods

et al. 2020

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“…In this table, we do not quote data on sensitivity that varies from 45% to 100% or specificity, almost always superior to 90%; heterogeneity of the methodology related to sample origins (vertebrate and invertebrate hosts, biological source of samples, etc. ), processing of samples (extraction of DNA with the various kit, other methodologies…), type of isothermal amplification and their targets, and downstream methods used to visualize results (Eyes, UV, colorimetric, agarose gel electrophoresis, oligo chromatography, fluorimetry, dedicated device…) are reviewed for Leishmania infection [ 35 , 36 ]. We instead analyze the steady-state level of tests to be translated into the field medical setting for visceral and cutaneous leishmaniasis, Chagas disease, and human or animal trypanosomosis.…”

Section: Field Detection Of Infections Due To Trypanosomatidae: Chall...mentioning

confidence: 99%

“…. ) are reviewed for Leishmania infection [35,36]. We instead analyze the steadystate level of tests to be translated into the field medical setting for visceral and cutaneous leishmaniasis, Chagas disease, and human or animal trypanosomosis.…”

Section: Does Isothermal Dna Amplification Represent the Ideal Tool F...mentioning

confidence: 99%

Isothermal Nucleic Acid Amplification to Detect Infection Caused by Parasites of the Trypanosomatidae Family: A Literature Review and Opinion on the Laboratory to Field Applicability

Sereno

Oury

Geiger

et al. 2022

IJMS

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Isothermal amplification of nucleic acids has the potential to be applied in resource-limited areas for the detection of infectious agents, as it does not require complex nucleic purification steps or specific and expensive equipment and reagents to perform the reaction and read the result. Since human and animal infections by pathogens of the Tryponasomatidae family occur mainly in resource-limited areas with scant health infrastructures and personnel, detecting infections by these methodologies would hold great promise. Here, we conduct a narrative review of the literature on the application of isothermal nucleic acid amplification for Trypanosoma and Leishmania infections, which are a scourge for human health and food security. We highlight gaps and propose ways to improve them to translate these powerful technologies into real-world field applications for neglected human and animal diseases caused by Trypanosomatidae.

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“…Several studies have shown LAMP as a useful tool for the rapid detection of bacterial, parasitic and viral pathogenic agents of infectious diseases including assays for 16 neglected tropical diseases such as leishmaniasis, schistosomiasis, Buruli ulcer, helminthic diseases or yaws [ 31 – 39 ]. Besides a range of in-house assays, commercial LAMP diagnostic kits developed by Eiken (Eiken Chemical Co., Tokyo, Japan) for human African trypanosomiasis (HAT) [ 40 ], tuberculosis (TB) [ 41 ], malaria [ 42 – 44 ], and leishmaniasis [ 45 , 46 ], as well as the centrifugation-free Illumigene assay for malaria genus-level detection developed by Meridian Biosciences (Cincinnati, USA) are available [ 47 , 48 ].…”

Section: Introductionmentioning

confidence: 99%

RLEP LAMP for the laboratory confirmation of leprosy: towards a point-of-care test

et al. 2021

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Background Nucleic acid-based amplification tests (NAAT), above all (q)PCR, have been applied for the detection of Mycobacterium leprae in leprosy cases and household contacts with subclinical infection. However, their application in the field poses a range of technical challenges. Loop-mediated isothermal amplification (LAMP), as a promising point-of-care NAAT does not require sophisticated laboratory equipment, is easy to perform, and is applicable for decentralized diagnosis at the primary health care level. Among a range of gene targets, the M. leprae specific repetitive element RLEP is regarded as highly sensitive and specific for diagnostic applications. Methods Our group developed and validated a dry-reagent-based (DRB) RLEP LAMP, provided product specifications for customization of a ready-to-use kit (intended for commercial production) and compared it against the in-house prototype. The assays were optimized for application on a Genie® III portable fluorometer. For technical validation, 40 “must not detect RLEP” samples derived from RLEP qPCR negative exposed and non-exposed individuals, as well as from patients with other conditions and a set of closely related mycobacterial cultures, were tested together with 25 “must detect RLEP” samples derived from qPCR confirmed leprosy patients. For clinical validation, 150 RLEP qPCR tested samples were analyzed, consisting of the following categories: high-positive samples of multibacillary (MB) leprosy patients (> 10.000 bacilli/extract), medium-positive samples of MB leprosy patients (1.001–10.000 bacilli/extract), low-positive samples of MB leprosy patients (1–1.000 bacilli/extract), endemic controls and healthy non-exposed controls; each n = 30. Results Technical validation: both LAMP formats had a limit of detection of 1.000 RLEP copies, i.e. 43–27 bacilli, a sensitivity of 92% (in-house protocol)/100% (ready-to-use protocol) and a specificity of 100%. Reagents were stable for at least 1 year at 22 °C. Clinical validation: Both formats showed a negativity rate of 100% and a positivity rate of 100% for high-positive samples and 93–100% for medium positive samples, together with a positive predictive value of 100% and semi-quantitative results. The positivity rate for low-positive samples was 77% (in-house protocol)/43% (ready-to-use protocol) and differed significantly between both formats. Conclusions The ready-to-use RLEP DRB LAMP assay constitutes an ASSURED test ready for field-based evaluation trials aiming for routine diagnosis of leprosy at the primary health care level.

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Loop-mediated isothermal amplification methods for diagnosis of visceral leishmaniasis (kala-azar) – a systematic review

Cited by 9 publications

References 35 publications

Laboratory Diagnosis of Cutaneous and Visceral Leishmaniasis: Current and Future Methods

Laboratory Diagnosis of Cutaneous and Visceral Leishmaniasis: Current and Future Methods

Isothermal Nucleic Acid Amplification to Detect Infection Caused by Parasites of the Trypanosomatidae Family: A Literature Review and Opinion on the Laboratory to Field Applicability

RLEP LAMP for the laboratory confirmation of leprosy: towards a point-of-care test

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