The Direct Boil-LAMP method: A simple and rapid diagnostic method for cutaneous leishmaniasis

Mikita, Kei; Maeda, Takeshi; Yoshikawa, Sachio; Ono, Takeshi; Miyahira, Yasushi; Kawana, Akihiko

doi:10.1016/j.parint.2014.07.007

Cited by 42 publications

(51 citation statements)

References 29 publications

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“…Compared with other NAA techniques such as PCR, the LAMP assay is more resistant to contamination with reaction-inhibitory substances often present in clinical samples. Since a large amount of contaminating substances inhibits the LAMP reaction, we previously evaluated the ability of PURE to eliminate the substrates that inhibit the LAMP reaction, including whole blood and debris from swab samples of a skin ulcer (10). We found that the PURE kit could be used to extract DNA from these clinical sample types and adequately eliminate the inhibitory substances, making it easy to detect DNA by combining it with the LAMP reaction.…”

Section: Discussionmentioning

confidence: 99%

PURE-LAMP Procedure for the Diagnosis of Extrapulmonary Tuberculosis: A Case Series

et al. 2015

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Although the polymerase chain reaction is effective for the diagnosis of extrapulmonary tuberculosis (EPTB), it is typically unavailable in resource-limited situations. In contrast, the loop-mediated isothermal amplification (LAMP) assay is a relatively cost-effective and accessible method. Additionally, when combined with the procedure for ultra-rapid extraction (PURE) kit, which enables simple DNA extraction, LAMP can detect Mycobacterium tuberculosis in sputum within 1.5 hours using a simple procedure. In this study, we investigated the utility of the PURE-LAMP technique to diagnose three cases of EPTB and showed that it may potentially be a valuable tool for the diagnosis of EPTB.

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Section: Discussionmentioning

confidence: 99%

PURE-LAMP Procedure for the Diagnosis of Extrapulmonary Tuberculosis: A Case Series

et al. 2015

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“…We also saw positive RT-LAMP reactions using DENV-positive samples that had been heat treated for 10 minutes at 100°C. Boiling samples can eliminate the need for formal RNA extraction steps as has been previously reported with DNA-based pathogen targets (Mikita et al, 2014;Hopkins et al, 2013).…”

Section: Assay Characteristicsmentioning

confidence: 96%

Development of a pan-serotype reverse transcription loop-mediated isothermal amplification assay for the detection of dengue virus

Dauner

Mitra

Gilliland

et al. 2015

Diagnostic Microbiology and Infectious Disease

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During dengue outbreaks, acute diagnosis at the patient's point of need followed by appropriate supportive therapy reduces morbidity and mortality. To facilitate needed diagnosis, we developed and optimized a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay that detects all 4 serotypes of dengue virus (DENV). We used a quencher to reduce nonspecific amplification. The assay does not require expensive thermocyclers, utilizing a simple water bath to maintain the reaction at 63 °C. Results can be visualized using UV fluorescence, handheld readers, or lateral flow immunochromatographic tests. We report a sensitivity of 86.3% (95% confidence interval [CI], 72.7-94.8%) and specificity of 93.0% (95% CI, 83.0-98.1%) using a panel of clinical specimens characterized by DENV quantitative reverse transcription-polymerase chain reaction. This pan-serotype DENV RT-LAMP can be adapted to field-expedient formats where it can provide actionable diagnosis near the patient's point of need.

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“…8 It has a high degree of sensitivity owing to the design and quantity of the primers used (i.e., two external primers, two internal primers, and two first loop primers). 10 Some studies have demonstrated the use of LAMP against extracts from macerated sandflies 8,11 and several alternatives exist for the visualization of the results including turbidity, fluorescence, and/or color changes. [8][9][10][12][13][14] To date, LAMP has been used for the diagnosis of bacterial, 15 fungal, 16,17 and viral 18,19 infections, as well as parasitic infections such as those caused by Trypanosoma brucei, 20 Plasmodium falciparum, 21 and Trypanosoma cruzi.…”

Section: Introductionmentioning

confidence: 99%

Analytical Performance of a Loop-Mediated Isothermal Amplification Assay for Leishmania DNA Detection in Sandflies and Direct Smears of Patients with Cutaneous Leishmaniasis

León

Muñoz

Tabares

et al. 2018

The American Journal of Tropical Medicine and Hygiene

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Loop-mediated isothermal amplification (LAMP) is ideal for the detection of DNA as it is a quick and easy-to-perform test that does not require complex or sophisticated equipment or infrastructure. However, the application of this technique in the detection of DNA has not been comprehensively analyzed to date (analytical validation). Our objective was to evaluate the sensitivity and analytical specificity (anticipated reportable range [ARR], the limit of detection [LoD], and accuracy) of LAMP targeting the 18S rRNA gene in the diagnosis of six New World species. We then applied the validated LAMP assay across 50 samples of sandflies and 50 direct smears from a recent outbreak of cutaneous leishmaniasis in Colombia to determine its diagnostic performance. The LAMP assay exclusively amplified the DNA of spp., and an ARR of between 1 × 10 and 1 × 10 equivalent parasites/mL was determined. An LoD of 1 × 10 equivalent parasites/mL was established and there was no statistically significant variation in terms of accuracy. Finally, a sensitivity of 100% in direct smears and sandflies samples was calculated and a specificity of 90.9% for direct smears using microscopy as reference and 96.8% for sandflies using real-time polymerase chain reaction as reference were determined. To our knowledge, this is the first attempt to analytically validate a LAMP test to detect DNA, which showed good diagnostic potential from sandflies and direct smear samples.

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The Direct Boil-LAMP method: A simple and rapid diagnostic method for cutaneous leishmaniasis

Cited by 42 publications

References 29 publications

PURE-LAMP Procedure for the Diagnosis of Extrapulmonary Tuberculosis: A Case Series

PURE-LAMP Procedure for the Diagnosis of Extrapulmonary Tuberculosis: A Case Series

Development of a pan-serotype reverse transcription loop-mediated isothermal amplification assay for the detection of dengue virus

Analytical Performance of a Loop-Mediated Isothermal Amplification Assay for Leishmania DNA Detection in Sandflies and Direct Smears of Patients with Cutaneous Leishmaniasis

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