Evaluation of real-time PCR for Strongyloides stercoralis and hookworm as diagnostic tool in asymptomatic schoolchildren in Cambodia

Schär, Fabian; Odermatt, Peter; Khieu, Virak; Panning, Marcus; Socheat, Duong; Muth, Sinuon; Marti, Hanspeter; Kramme, Stefanie

doi:10.1016/j.actatropica.2012.12.012

Cited by 89 publications

(101 citation statements)

References 17 publications

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“…Whilst real-time PCR has shown very high specificity in human samples, it again has limited sensitivity, and has not proven to be diagnostically superior to other parasitologic techniques such as the Baermann method or APC [61,63,64•], particularly in low-density infections, supporting previous findings from animal models [65,66]. However, recent improvements of this technique may increase the sensitivity in the near future.…”

Section: Molecular Biologysupporting

confidence: 48%

Advances in the Diagnosis of Human Strongyloidiasis

et al. 2014

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Strongyloidiasis is an intestinal parasitic infection that is particularly relevant in immunosuppressed patients because it can cause severe disseminated disease. This review discusses the recent advances in the diagnosis of strongyloidiasis. We suggest clinical and epidemiologic criteria for the diagnosis and screening of strongyloidiasis, taking into account different epidemiologic contexts. The state of the art of the diagnosis of strongyloidiasis is discussed including parasitologic methods that are commonly used despite having low sensitivity; serology, which has demonstrated better sensitivity (with some exceptions such as travelers or immunosuppressed patients), and molecular biology methods, which have virtually 100 % specificity. Finally, we discuss different strategies to follow up patients after treatment, highlighting the importance of having accurate and reliable follow-up markers when assessing treatment efficacy both in a clinical and research context.

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Section: Molecular Biologysupporting

confidence: 48%

Advances in the Diagnosis of Human Strongyloidiasis

et al. 2014

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“…Además se pueden producir reacciones cruzadas con otros helmintos y no permiten diferenciar entre las DIAGNÓSTICO DE LA INFECCIÓN POR STRONGYLOIDES STERCORALIS: METANÁLISIS SOBRE EVALUACIÓN DE MÉTODOS ... infecciones actuales y las pasadas 2 . Igualmente, la reacción en cadena de la polimerasa (PCR) también se ha utilizado como método diagnóstico, presentando diferencias en la sensibilidad obtenida en diferentes ensayos [27][28][29] .…”

Section: Introductionunclassified

Infección por Strongyloides stercoralis: metanálisis sobre evaluación de métodos diagnósticos convencionales (1980-2013)

Campo-Polanco

Gutiérrez

Arias

2014

Rev. Esp. Salud Publica

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(*) El presente trabajo es un producto derivado del desarrollo de la propuesta de investigación "Diagnós-tico molecular de Estrongiloidiasis mediante PCR en tiempo rea. Comparación con métodos de diagnós-tico tradicionales" aprobado por el Centro de Investigación para el Desarrollo y la Innovación (CIDI), Universidad Pontificia Bolivariana. Medellín. Colombia en la convocatoria XIV. código interno de radicado 175-11/11 935A. Sin conflicto de intereses RESUMENFundamentos: Las investigaciones sobre los métodos diagnósticos de estrongiloidiasis presentan divergencias en su validez y son incompletas al no reportar datos sobre seguridad, eficiencia y desempeño diagnós-tico. El objetivo del trabajo fue evaluar la validez, desempeño, eficiencia y seguridad diagnósticas de cuatro métodos parasitológicos convencionales para la detección de la infección por Strongyloides stercoralis. Métodos:Revisión sistemática con metanálisis, búsqueda exhaustiva y reproducible de literatura en seis bases de datos para el período 1980-2013. Se evaluó la calidad de los artículos y se realizó metanálisis con el modelo de efectos aleatorios, calculando sensibilidad, especificidad, cocientes de probabilidad, valores predictivos, proporción de resultados falsos, exactitud, razón de odds, índice J de Youden y curva ROC, usando Meta-DiSc(es) y Epidat 3.1.Resultados: Se incluyeron 11 estudios con 9.025 individuos. La sensibilidad del método de Baermann fue del 72%, cociente de probabilidad positivo (CP+) 228 y cociente de probabilidad negativo (CP-) 0,32. El en placa de ágar (CPA) presentó una sensibilidad de 89%, CP+ 341 y CP-0,11. En el examen directo en heces la sensibilidad fue del 21%; el CP+ 67 y el CP-0,67. La sensibilidad de la concentración formol-éter fue del 48%, el CP+ 110 y el CP-0,59. Áreas bajo la curva ROC de 0,999 en Baermann y CPA; 0,977 en el examen directo en heces y 0,829 en concentración formol-éter; la especificidad fue de 100% en todas las pruebas. Conclusiones:Los cuatro métodos parasitológicos convencionales evaluados en este estudio para la detección de S. stercoralis pueden ser útiles, sin embargo, resulta más idóneo implementar el en placa de ágar y el método de Baermann.

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“…8,19,37 Some of this variation in sensitivity may be caused by the use of different extraction techniques and the period of time stool specimens were stored frozen or in ethanol, before DNA extraction. 8,19,37 The comparison of the LAMP assay to PCR performed on extracts from stool that were microscopy positive for S. stercoralis showed concordance for samples with a wide range of PCR C t values. The only sample that was negative using the LAMP assay had a high C t value ( 38) with real-time PCR.…”

Section: Discussionmentioning

confidence: 99%

A Loop-Mediated Isothermal Amplification (LAMP) Assay for Strongyloides stercoralis in Stool That Uses a Visual Detection Method with SYTO-82 Fluorescent Dye

Watts

James

Sultana

et al. 2014

The American Society of Tropical Medicine and Hygiene

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Abstract. An assay to detect Strongyloides stercoralis in stool specimens was developed using the loop-mediated isothermal amplification (LAMP) method. Primers were based on the 28S ribosomal subunit gene. The reaction conditions were optimized and SYTO-82 fluorescent dye was used to allow real-time and visual detection of the product. The product identity was confirmed with restriction enzyme digestion, cloning, and sequence analysis. The assay was specific when tested against DNA from bacteria, fungi and parasites, and 30 normal stool samples. Analytical sensitivity was to 10 copies of target sequence in a plasmid and up to a 10 -2 dilution of DNA extracted from a Strongyloides ratti larva spiked into stool. Sensitivity was increased when further dilutions were made in water, indicative of reduced reaction inhibition. Twenty-seven of 28 stool samples microscopy and polymerase chain reaction positive for S. stercoralis were positive with the LAMP method. On the basis of these findings, the assay warrants further clinical validation.

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Evaluation of real-time PCR for Strongyloides stercoralis and hookworm as diagnostic tool in asymptomatic schoolchildren in Cambodia

Cited by 89 publications

References 17 publications

Advances in the Diagnosis of Human Strongyloidiasis

Advances in the Diagnosis of Human Strongyloidiasis

Infección por Strongyloides stercoralis: metanálisis sobre evaluación de métodos diagnósticos convencionales (1980-2013)

A Loop-Mediated Isothermal Amplification (LAMP) Assay for Strongyloides stercoralis in Stool That Uses a Visual Detection Method with SYTO-82 Fluorescent Dye

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