Molecular Diagnosis of Polycystic Echinococcosis Due to Echinococcus vogeli in a Paraguayan Immigrant in Argentina

Grenouillet, Frédéric; Frider, Bernardo; Rodríguez, Juan Alvarez; Amante, Marcelo; Pestalardo, María Luján; Cazorla, Arnault; Bresson‐Hadni, Solange; Millon, Laurence

doi:10.1128/jcm.00871-13

Cited by 20 publications

(5 citation statements)

References 18 publications

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“…The most likely reason for this is a loss of DNA stability in conserved FFPE material, as has been previously described [ 38 ]. It has already been suggested that in such cases the application of shorter amplicons might lead to higher resolution [ 39 ] but at least in our case, the utilization of an amplicon of around 200 bp did not yield improved results. This does not exclude, however, that the utilization of alternative primers, shorter amplicons, or specific DNA purification methodology for FFPE material (e.g.…”

Section: Discussioncontrasting

confidence: 56%

“…Since DNA degradation in FFPE samples of longer storage is most probably due to DNA fragmentation [ 38 ], the utilization of shorter amplicons up to 200 bp has been suggested as possible improvement [ 39 ]. We therefore designed another primer (12S-newR2) which, together with primer 12S-newF should amplify a fragment of 287 nt and tested this combination on isolated parasite cells mixed with 25 mg liver tissue of M .…”

Section: Resultsmentioning

confidence: 99%

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Establishing and evaluation of a polymerase chain reaction for the detection of Echinococcus multilocularis in human tissue

Grimm

Krickl²,

Beck

et al. 2021

PLoS Negl Trop Dis

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Background Alveolar echinococcosis (AE) is caused by metacestode larva of the tapeworm Echinococcus multilocularis. AE diagnostics currently rely on imaging techniques supported by serology, but unequivocal detection of AE is difficult. Although polymerase chain reaction (PCR)-based methods to detect tapeworm DNA in biopsies have been suggested for several species, no validated protocol adhering to accepted guidelines has so far been presented for AE diagnostics. We herein established a PCR protocol for metacestode biopsies and technically evaluated the method using isolated parasite DNA and cells, biopsies of clinically relevant material, and formalin fixed paraffin-embedded (FFPE) human tissue blocks. We compared the results with an immunochemical (IHC) approach using the monoclonal antibody Em2G11 specific for the antigen Em2 of E. mulitlocularis. Methodology/Principal findings Based on tapeworm 12S rDNA sequences we established and validated a PCR protocol for robust detection of as little as 50 parasite cells per specimen and report 127 cases of positive identification of Echinococcus species in samples from humans and animals. For further validation, we analyzed 45 liver, heart, brain, and soft tissue samples as well as cytological probes of aspirates of FFPE-material from 18 patients with clinically confirmed AE. Of each patient we analyzed (i) fully viable lesions with laminated layer; (ii) tissue with mAbEm2G11-positive small particles of E. multilocularis (spems); (iii) mAbEm2G11-negative tissue adjacent to the main lesion; and (iv) lymph node tissue with mAbEm2G11-positive spems. To identify the areas for the PCR-based approach, we performed IHC-staining with the monoclonal antibody Em2G11. Micro-dissected tissue of these areas was then used for PCR-analysis. 9 of 15 analyzed samples with viable E. multilocularis lesions with laminated layer were positive by PCR. Of this group, all samples preserved for less than 6 years (6/6) were tested positive. 11 of 15 samples of spems and 7 of 9 samples of the control group mAbEm2G11-negative tissue were negative by PCR. We further show that all probes from lymph nodes with spems are PCR negative. Conclusions/Significance We present a sensitive PCR method for the detection of E. multilocularis in human tissue, particularly in fresh biopsy material and tissue blocks stored for less than 5 years. While the diagnostic sensitivity of material containing only spems was higher using IHC, PCR detection was possible in IHC negative liver tissue and in patients with negative serology. Our results support the view that spems do not contain parasitic DNA or viable cells of the parasite. spems thus most probably do not directly contribute to metastasis formation during AE.

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Section: Discussioncontrasting

confidence: 56%

Section: Resultsmentioning

confidence: 99%

Establishing and evaluation of a polymerase chain reaction for the detection of Echinococcus multilocularis in human tissue

Grimm

Krickl²,

Beck

et al. 2021

PLoS Negl Trop Dis

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“…(4) Another human case of infection but by E. vogeli was reported from Argentina, the female patient was a Paraguayan immigrant who presented jaundice and abdominal pain in the right hypochondriac region. The diagnosis was achieved by abdominal CT, immunodiagnosis techniques (ELISA, immunoelectophoresis, Western blot), histopathologic studies, and molecular techniques (PCR) [37]. On the contrary, a review was recently published on the genus Echinococcus spp.…”

Section: Resultsmentioning

confidence: 99%

Neotropical Echinococcosis: A Review

Meléndez¹

2022

Zoonosis of Public Health Interest

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Echinococcus vogeli (Rausch and Berstein, 1972) and Echinococcus oligarthra (Diesing, 1863) (Cestoda: Taeniidae) are the only two species known of Neotropical tapeworms, which cause Echinococcosis Polycystic (EP) and Echinococcosis Unicystic (EU), respectively, in humans and in wild rodents from Central and South America. This review applied a meta-analysis on published research about these diseases during the last decade (2010–2020) with the aim of finding out the new human cases reported on that decade on EP and EU. Several new human cases have been published in these 10 years, and important findings have been carried out on the phylogenetic taxonomy, on the genome of E. oligarthra, and on new molecular diagnostic techniques and imagenology applied upon this two neotropical echinococcosis, in particular in Argentina and Brazil. Finally, the life cycle of both Echinococcus species appears to be in a dynamic activity, apparently there is an expansion of both zoonotic diseases moving down to Southern zones of Argentina; therefore, a program of epidemiological surveillance on EP and EU is proposed to be carried out in those Patagonic regions.

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“…Indeed, the amplification of the above-mentioned sample SI-H4 at the rrnS locus was successful only when a shorter fragment of this gene was amplified. However, as discussed in previous studies, the use of a target size shorter than 200 bp might have increased the sensitivity even further [ 52 , 53 ]. An interesting approach was described in the study of Koonmee et al [ 52 ], where repeated PCR using the same primer sets and conditions was successfully employed for the amplification of short DNA fragments from formalin-fixed paraffin-embedded (FFPE) tissues.…”

Section: Discussionmentioning

confidence: 99%

Molecular Characterization of Echinococcus granulosus sensu lato from Humans in Slovenia

et al. 2020

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The larval form of tapeworms of the Echinococcus granulosus sensu lato species cluster cause an important zoonotic infection, cystic echinococcosis (CE). Molecular characterization of the cluster’s isolates from different hosts greatly contributes to a better understanding of its transmission dynamics. To date, no genetic information is available on CE in Slovenia. In this work, we characterized isolates from human CE cases. Parasite samples from 18 patients were collected, together with the patients’ demographic and clinical data. Genomic DNA was analyzed by conventional PCR and sequencing at four mitochondrial loci (cytochrome c oxidase subunit 1, cox1; NADH dehydrogenase subunit 1, nad1; NADH dehydrogenase subunit 5, nad5; and small ribosomal RNA, rrnS). Thirteen isolates were successfully amplified and sequenced. Seven (58.8%) patients were infected with E. granulosus sensu stricto (s.s.) G1, five (38.5%) with E. canadensis G7 and one (7.7%) with E. granulosus s.s. G3. Echinococcus canadensis G7, the pig genotype, was identified exclusively in autochthonous Slovenes, while the patients originating from the Western Balkans were all infected with E. granulosus s.s. Our findings suggest that pigs are important intermediate hosts for human CE in Slovenia.

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Molecular Diagnosis of Polycystic Echinococcosis Due to Echinococcus vogeli in a Paraguayan Immigrant in Argentina

Cited by 20 publications

References 18 publications

Establishing and evaluation of a polymerase chain reaction for the detection of Echinococcus multilocularis in human tissue

Establishing and evaluation of a polymerase chain reaction for the detection of Echinococcus multilocularis in human tissue

Neotropical Echinococcosis: A Review

Molecular Characterization of Echinococcus granulosus sensu lato from Humans in Slovenia

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