Real-Time Polymerase Chain Reaction for Detection of Strongyloides stercoralis in Stool

Sultana, Yasmin; Jeoffreys, Neisha; Watts, Matthew; Gilbert, Gwendolyn L.; Lee, Rogan

doi:10.4269/ajtmh.12-0437

Cited by 64 publications

(77 citation statements)

References 19 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: 46%

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: 46%

Advances in the Diagnosis of Human Strongyloidiasis

et al. 2014

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“…Analytical sensitivity and specificity are comparable to PCR, according to the method of Verweij et al 19,20,23 .…”

Section: Pcrmentioning

confidence: 99%

“…No PCR studies have reported false positive results when analytical specificity has been tested using DNA extracted from bacteria, viruses, fungi, protozoa, and other helminths 19,23,24,27,29,30 . Studies have also assessed the specificity of the PCR products by sequence analysis, with all finding 100% sequence homology with the target sequence of S. stercoralis.…”

Section: Pcrmentioning

confidence: 99%

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The laboratory diagnosis of Strongyloides stercoralis

Watts¹,

Robertson²,

Bradbury

2016

Microbiol. Aust.

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It is estimated that over 30million people worldwide are infected by the nematode, Strongyloides stercoralis1. It is endemic in sub-tropical and tropical parts of Australia, with high rates of infection documented in some indigenous communities2. Due to the potential for chronic autoinfection, that may persist for decades, migration leads to the presence of the infection in non-endemic areas1. Transmission to humans is generally through the penetration of larvae through the skin, following contact with faecally contaminated soil1. Disease severity ranges from asymptomatic chronic carriage to an overwhelming illness, where large numbers spread throughout the body, usually triggered by immunosuppression1.

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“…18 The DNA was extracted from S. ratti larvae in water and larvae spiked into normal human stool using a spin column method, the PowerSoil DNA Isolation Kit (Mo Bio Laboratories, Carlsbad CA), according to the manufacturer's instructions. 19 The DNA was extracted from a range of other parasites, fungi, and bacteria using standard methods (Table 1). These samples were either from pure cultures or, in the case of parasites, washed and concentrated organisms.…”

Section: Methodsmentioning

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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Real-Time Polymerase Chain Reaction for Detection of Strongyloides stercoralis in Stool

Cited by 64 publications

References 19 publications

Advances in the Diagnosis of Human Strongyloidiasis

Advances in the Diagnosis of Human Strongyloidiasis

The laboratory diagnosis of Strongyloides stercoralis

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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