Diagnóstico de las infecciones por geohelmintos. Un problema sin resolver en la era de las ómicas

Fernández‐Rivas, Gema; Rivaya, Belén; Romaní, Nona; Wang, Jun Hao; Alcaide, Mireya; Matas, Lurdes

doi:10.1016/s0213-005x(19)30178-8

Cited by 3 publications

(2 citation statements)

References 17 publications

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“…Until now, few recombinant proteins have been studied as potential immunogenic molecules for serodiagnosis of strongyloidiasis [16,44]. The antibodies against rSs14-3-3 and rSsMA are not increased with the immunization and they then do not provide any protective effect, but their quantification could be useful for diagnosis and as a marker of the severity of the infection [44,45].…”

Section: Discussionmentioning

confidence: 99%

Evaluation of the Immunomodulatory Effect of the Recombinant 14-3-3 and Major Antigen Proteins of Strongyloides stercoralis against an Infection by S. venezuelensis

et al. 2022

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Strongyloidiasis, caused by Strongyloides stercoralis, is a neglected parasitic disease that represents a serious public health problem. In immunocompromised patients, this parasitosis can result in hyperinfection or disseminated disease with high levels of mortality. In previous studies, the mRNAs encoding for the 14-3-3 and major antigen proteins were found to be expressed at high levels in S. stercoralis L3 larvae, suggesting potential key roles in parasite-host interactions. We have produced them as recombinant proteins (rSs14-3-3 and rSsMA) in a bacterial protein expression system. The serum levels of anti-rSs14-3-3 and anti-rSsMA IgGs are increased upon infection with S. venezuelensis, validating the use of the mouse model since the native 14-3-3 and MA proteins induce an immune response. Each recombinant protein was formulated in the adjuvant adaptation (ADAD) vaccination system and injected twice, subcutaneously, in CD1 mice that were experimentally infected with 3000 S. venezuelensis L3 to evaluate their protective and immunomodulatory activity. Our results, including the number of parthenogenetic females, number of eggs in stool samples and the analysis of the splenic and intestinal indexes, show that the vaccines did not protect against infection. The immunization with rSs14-3-3 induced changes in the cytokine profile in mice, producing higher expression of IL-10, TGF-β, IL-13 and TNF-α in the spleen, suggesting a Th2/Treg-type response with an increase in TNF-α levels, confirming its role as an immunomodulator.

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Section: Discussionmentioning

confidence: 99%

Evaluation of the Immunomodulatory Effect of the Recombinant 14-3-3 and Major Antigen Proteins of Strongyloides stercoralis against an Infection by S. venezuelensis

et al. 2022

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“…There is much controversy as to whether S. stercoralis represents a zoonosis acquired from dogs and cats. In the last two decades, various molecular techniques have been applied to Patients infected with S. stercoralis may have low parasite loads, so the low sensitivity of parasitological methods and the low specificity of immunological methods, due to cross-reactions with other helminths, do not allow an adequate diagnosis of the disease (Buonfrate et al 2015a(Buonfrate et al , 2015bFernández-Rivas et al 2016).…”

Section: Conditions Tested For Amplification Of S Stercoralis Sequenc...mentioning

confidence: 99%

Comparison of different PCR amplification targets for molecular diagnosis of Strongyloides stercoralis

Marquet,

Mora,

Incani

et al. 2023

J. Helminthol.

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Molecular techniques are an alternative for the diagnosis of strongyloidiasis, produced by Strongyloides stercoralis. However, it is necessary to determine the best amplification target for the populations of this parasite present in a geographical area and standardize a polymerase chain reaction (PCR) protocol for its detection. The objectives of this work were the comparison of different PCR targets for molecular detection of S. stercoralis and the standardization of a PCR protocol for the selected target with the best diagnostic results. DNA extraction was performed from parasite larvae by saline precipitation. Three amplification targets of the genes encoding ribosomal RNA 18S (18S rDNA) and 5.8S (5.8S rDNA) and cytochrome oxidase 1 (COX1) of S. stercoralis were compared, and the PCR reaction conditions for the best target were standardized (concentration of reagents and template DNA, hybridization temperature, and number of cycles). The analytical sensitivity and specificity of the technique were determined. DNA extraction by saline precipitation made it possible to obtain DNA of high purity and integrity. The ideal target was the 5.8S rDNA, since the 18S rDNA yielded non-reproducible results and COX1 never amplified under any condition tested. The optimal conditions for the 5.8S rDNA-PCR were: 1.5 mM MgCl2, 100 μM dNTPs, 0.4 μM primers, and 0.75 U DNA polymerase, using 35 cycles and a hybridization temperature of 60 °C. The analytical sensitivity of the PCR was 1 attogram of DNA, and the specificity was 100%. Consequently, the 5.8S rDNA was shown to be highly sensitive and specific for the detection of S. stercoralis DNA.

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Parasitosis con manifestaciones digestivas

López¹,

Bueno²,

Leiro³

et al. 2022

Medicine - Programa de Formación Médica Continuada Acreditado

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Diagnóstico de las infecciones por geohelmintos. Un problema sin resolver en la era de las ómicas

Cited by 3 publications

References 17 publications

Evaluation of the Immunomodulatory Effect of the Recombinant 14-3-3 and Major Antigen Proteins of Strongyloides stercoralis against an Infection by S. venezuelensis

Evaluation of the Immunomodulatory Effect of the Recombinant 14-3-3 and Major Antigen Proteins of Strongyloides stercoralis against an Infection by S. venezuelensis

Comparison of different PCR amplification targets for molecular diagnosis of Strongyloides stercoralis

Parasitosis con manifestaciones digestivas

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