Development of a low-cost copro-LAMP assay for simultaneous copro-detection of <i>Toxocara canis</i> and <i>Toxocara cati</i>

Avila, Héctor Gabriel; Risso, Marikena Guadalupe; Ruybal, Paula; Repetto, Silvia; Butti, Marcos Javier; Trangoni, Marcos David; Löffler, Sylvia Grune; Pérez, Verónica Mirtha; Periago, María Victoria

doi:10.1017/s0031182021000342

Cited by 8 publications

(9 citation statements)

References 63 publications

(79 reference statements)

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“…The Copro-LAMPAc was able to detect 100 fg of gDNA from A. caninum adult worms, which is more sensitive than previously reported values for other PCR assays (33)(34)(35)66). This analytical sensitivity value is included in the femtograms range, similar to what has been obtained in other studies (38,49,74). In future studies, the A. caninum egg limit of detection could be determined.…”

Section: Discussionsupporting

confidence: 79%

“…The DNA integrity and concentrations were determined according to Avila et al ( 38 ). The DNA extraction and purification from feces (fDNA) was obtained using a commercial kit (CKM) and an alternative low-cost method (LCM) based on mesh-filtration of stool, followed by alkaline hydrolyses, according to Avila et al ( 38 ).…”

Section: Methodsmentioning

confidence: 99%

“…The selection of the target gene for the primer design was performed according to Avila et al ( 38 ), using Primer V5 design software ( 61 ). The target for primer design was a 208 bp region of the mitochondrial gene cox -1 (Genbank accession number NC_012309.1 , region: 293-501 bp).…”

Section: Methodsmentioning

confidence: 99%

“…Different protocols based on LAMP reactions, have been implemented for diagnosis of different parasites (30,38,(40)(41)(42)(43)(44)(45)(46)(47)(48)(49). Similarly, LAMP reactions have been shown to be useful for the detection of pathogens in food and surveillance of water quality (50)(51)(52)(53)(54)(55)(56)(57).…”

Section: Introductionmentioning

confidence: 99%

“…On the other hand, the isothermal amplification of nucleic acids has begun to be widely used since it can be carried out in laboratories without specialized equipment. Particularly, the LAMP (loop-mediated isothermal amplification) technique ( 37 ), uses 3 primer pairs that recognize a small DNA fragment, and generate looped structures that serve as a template to start a new polymerization cycle thus providing both higher specificity and sensitivity ( 38 ). The DNA polymerase I from Bacillus stearothermophilus ( Bst ) used in the technique causes DNA strand displacement and therefore does not require denaturing the double strand, thus the technique can be performed with any equipment that guarantees a constant temperature ( 37 ).…”

Section: Introductionmentioning

confidence: 99%

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Development of a New LAMP Assay for the Detection of Ancylostoma caninum DNA (Copro-LAMPAc) in Dog Fecal Samples

et al. 2021

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Ancylostoma caninum is a zoonotic nematode which is able to affect animals and humans. Diagnosis in the definitive host and environmental detection are key to prevent its dissemination and achieve control. Herein, a new coprological LAMP method for the detection of A. caninum (Copro-LAMPAc) DNA was developed. DNA extraction was performed using a low-cost method and a fragment of the cox-1 gene was used for primer design. The analytical sensitivity, evaluated with serial dilutions of genomic DNA from A. caninum adult worms, was 100 fg. A specificity of 100% was obtained using genomic DNA from the host and other pathogens. The Copro-LAMPAc was evaluated using environmental canine fecal samples. When compared with gold standard optical microscopy in epidemiological studies, it proved to be more sensitive. This new LAMP assay can provide an alternative protocol for screening and identification of A. caninum for epidemiological studies in endemic areas.

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