Loop-Mediated Isothermal Amplification Allows Rapid, Simple and Accurate Molecular Diagnosis of Human Cutaneous and Visceral Leishmaniasis Caused by Leishmania infantum When Compared to PCR

Ibarra-Meneses, Ana Victoria; Chicharro, Carmen; Sánchez, C.; García, E. Barbero; Ortega, Sergio; Ndung’u, Joseph Mathu; Moreno, Javier; Cruz, Israel; Carrillo, Eugenia

doi:10.3390/microorganisms9030610

Cited by 4 publications

(2 citation statements)

References 22 publications

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“…Además de las metodologías de qPCR, es también necesario establecer otras alternativas de diagnós tico molecular para la LC en Panamá, especial mente aquellas de fácil implementación en entornos de áreas endémicas. Este es el caso de la amplifi cación isotérmica mediada por bucle (LAMP), ya que no requiere de un termociclador, el tiempo total de amplificación es corto, la visualización de los re sultados puede ser directa, reporta altas sensibili dades y bajo riesgo de contaminación [40].…”

Section: Discussionunclassified

Comparación de pruebas moleculares para el diagnóstico de Leishmania spp. en lesiones cutáneas con baja carga parasitaria

Reina,

Pineda,

Calzada

et al. 2023

Rev Med Panama

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Introducción: La leishmaniasis cutánea (LC) es un problema grave de salud pública en Panamá. El diagnóstico de esta parasitosis ha sido siempre desafiante, no sólo debido a su similitud con otras infecciones dérmicas, sino también a características particulares de las lesiones, como cargas parasitarias bajas. Materiales y Métodos: En este estudio, se evaluaron mediante cuatro métodos moleculares, 235 muestras de ADN procedentes de lesiones con frotis negativos por LC obtenidas durante el período 2015-2019. Resultados: Los resultados señalan que las sensibilidades encontradas fueron de 75.6% (IC 0.6234-0.8709) para la PCR kDNA-Género específico, de 66.7% (IC 0.5359-0.776) para la PCR Hsp70-Género específico y de 77.6% (IC 0.645-0.8949) para la qPCR 18S ribosomal. Todas las pruebas obtuvieron un valor predictivo positivo de 100%, mientras que el valor predictivo negativo más alto fue con la qPCR (80.58%) y el más bajo con el PCR Hsp70-Género específico (73.2%). En cuanto a la precisión de diagnóstico se obtuvo un rango mayor del 82% en todas las pruebas evaluadas. Conclusión: Este estudio confirma la buena sensibilidad de la PCR kDNA-Viannia para el análisis de lesiones de LC con baja carga parasitaria. Esta metodología es relativamente fácil de estandarizar, por lo que se recomienda su uso en laboratorios clínicos regionales de Panamá. Aun cuando la qPCR 18S ribosomal presentó una sensibilidad relativamente menor, el uso de esta metodología debe ser también considerada por su facilidad de uso, menor tiempo de ejecución y capacidad de cuantificación.

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Section: Discussionunclassified

Comparación de pruebas moleculares para el diagnóstico de Leishmania spp. en lesiones cutáneas con baja carga parasitaria

Reina,

Pineda,

Calzada

et al. 2023

Rev Med Panama

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“…A real-time PCR assay has also been developed for A. baumannii detection [ 56 ], which provides faster results compared with conventional PCR. However, PCR assays require the use of expensive equipment and thus may not be an ideal solution for some countries [ 57 , 58 ]. Accurate and early detection of A. baumannii infections is very essential to efficient healthcare for patients due to the high mortality rate associated with its infections [ 59 ].…”

Section: Discussionmentioning

confidence: 99%

Sequence-Specific Electrochemical Genosensor for Rapid Detection of blaOXA-51-like Gene in Acinetobacter baumannii

et al. 2022

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Acinetobacter baumannii (A. baumannii) are phenotypically indistinguishable from the Acinetobacter calcoaceticus–A. baumannii (ACB) complex members using routine laboratory methods. Early diagnosis plays an important role in controlling A. baumannii infections and this could be assisted by the development of a rapid, yet sensitive diagnostic test. In this study, we developed an enzyme-based electrochemical genosensor for asymmetric PCR (aPCR) amplicon detection of the blaOXA-51-like gene in A. baumannii. A. baumannii blaOXA-51-like gene PCR primers were designed, having the reverse primer modified at the 5′ end with FAM. A blaOXA-51-like gene sequence-specific biotin labelled capture probe was designed and immobilized using a synthetic oligomer (FAM-labelled) deposited on the working electrode of a streptavidin-modified, screen-printed carbon electrode (SPCE). The zot gene was used as an internal control with biotin and FAM labelled as forward and reverse primers, respectively. The blaOXA-51-like gene was amplified using asymmetric PCR (aPCR) to generate single-stranded amplicons that were detected using the designed SPCE. The amperometric current response was detected with a peroxidase-conjugated, anti-fluorescein antibody. The assay was tested using reference and clinical A. baumannii strains and other nosocomial bacteria. The analytical sensitivity of the assay at the genomic level and bacterial cell level was 0.5 pg/mL (1.443 µA) and 103 CFU/mL, respectively. The assay was 100% specific and sensitive for A. baumannii. Based on accelerated stability performance, the developed genosensor was stable for 1.6 years when stored at 4 °C and up to 28 days at >25 °C. The developed electrochemical genosensor is specific and sensitive and could be useful for rapid, accurate diagnosis of A. baumannii infections even in temperate regions.

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Diagnostic performance of CL Detect rapid-immunochromatographic test for cutaneous leishmaniasis: a systematic review and meta-analysis

Gebremeskele,

Adane,

Adem

et al. 2023

Syst Rev

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Background Sensitive, robust, and fast point-of-care tests are needed for cutaneous leishmaniasis (CL) diagnosis. The recently developed CL Detect rapid test (InBios) for detecting Leishmania peroxidoxin antigen has been evaluated in several studies. However, diagnostic performances were controversial. Therefore, this systematic review and meta-analysis aimed to determine the pooled sensitivity and specificity of CL Detect for CL diagnosis. Methods PubMed, Scopus, EMBASE, ScienceDirect, and Google Scholar were sources of articles. We included studies reporting the diagnostic accuracy of CL Detect and CL-suspected patients in the English language. The methodological qualities of the included studies were appraised using the quality assessment of diagnostic accuracy studies-2 (QUADAS‐2). Meta-analysis was conducted using Stata 14.2 and R software. Results A total of 9 articles were included. The study sample size ranged from 11 to 274. The sensitivities of the individual studies ranged from 23 to 100%, and the specificities ranged from 78 to 100%. Pooled sensitivity and specificity were 68% (95% CI, 41–86%) and 94% (95% CI, 87–97%), respectively. AUC displayed 0.899. Pooled sensitivity was lower (47%, 95% CI, 34–61%) when PCR was used as a reference than microscopy (83%, 95% CI, 39–97%). Pooled sensitivity was lower (48%, 95% CI, 30–67%) for all lesion durations compared to ≤ 4 months (89%, 95% CI, 43–99%). Conclusions CL Detect has poor sensitivity and does not meet the minimal sensitivity of 95% of target product profiles designed for CL point-of-care tests. Currently, the CL Detect test looks unsuitable for CL diagnosis, despite its high specificity. Findings are limited by the low number of studies available. Further large-scale studies are recommended. Systematic review registration PROSPERO CRD42022323497.

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Loop-Mediated Isothermal Amplification Allows Rapid, Simple and Accurate Molecular Diagnosis of Human Cutaneous and Visceral Leishmaniasis Caused by Leishmania infantum When Compared to PCR

Cited by 4 publications

References 22 publications

Comparación de pruebas moleculares para el diagnóstico de Leishmania spp. en lesiones cutáneas con baja carga parasitaria

Comparación de pruebas moleculares para el diagnóstico de Leishmania spp. en lesiones cutáneas con baja carga parasitaria

Sequence-Specific Electrochemical Genosensor for Rapid Detection of blaOXA-51-like Gene in Acinetobacter baumannii

Diagnostic performance of CL Detect rapid-immunochromatographic test for cutaneous leishmaniasis: a systematic review and meta-analysis

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