A Multiplex-PCR approach to identification of the Brazilian intermediate hosts of Schistosoma mansoni

Vidigal, Teofânia H. D. A.; Magalhães, Kelly Grace; Kissinger, Jessica C.; Caldeira, Roberta Lima; Simpson, Andrew J.G.; Carvalho, Omar S.

doi:10.1590/s0074-02762002000900019

Cited by 9 publications

(6 citation statements)

References 14 publications

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“…In this respect, the ITS1 region seems to be more amenable to generate species specific SSCP patterns for this genus because of the presence of a higher number of nucleotide polymorphisms and deletions among the different Aplysina species, as illustrated in Figure 2. Interestingly, the ITS2 region has been described as being more variable than the ITS1 region for sponges (Wörheide et al , 2004) and other organisms (Deprés et al , 1995; Vidigal et al , 2000, 2002). These observations diverge from earlier data obtained by us in which the ITS1 region seemed to present higher variability than the ITS2 region for Hymeniacidon aff.…”

Section: Discussionmentioning

confidence: 99%

Aplysina (Porifera: Demospongiae) species identification through SSCP-ITS patterns

et al. 2009

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Keratose sponges have no spicules and limited robust morphological characters, causing severe problems for taxonomic classification. Single-strand conformational polymorphism (SSCP) analysis of PCR amplified internal transcribed spacers (ITS) of nuclear ribosomal RNA (rRNA) genes was used to elucidate speciation patterns in marine sponges of the genus Aplysina. In this study, Aplysina species with non-ambiguous traditional morphological alpha taxonomy were used to generate standard SSCP-ITS patterns, which were compared with patterns from alleged new species. Our results show that ITS-SSCP patterns allowed the differentiation of Aplysina sponges, demonstrating the advantage of a method that explores the secondary structure compared to direct sequencing.

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Section: Discussionmentioning

confidence: 99%

Aplysina (Porifera: Demospongiae) species identification through SSCP-ITS patterns

et al. 2009

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“…However, few authors developed speciesspecific primers for Biomphalaria spp., the intermediate host of S. mansoni. Vidigal et al (2002) designed specific polymerase chain reaction (PCR) primers for Brazilian snail hosts of S. mansoni from available sequences of internal transcribed spacer 2 (ITS2) of the ribosomal RNA gene. From the previous sequencing of the ITS2 region, one primer was designed to anchor in the 5.8S conserved region and three other species-specific primers in the 28S region, flanking the ITS2 region.…”

Section: Discussionmentioning

confidence: 99%

“…Vidigal et al (2002) designed specific PCR primers for Brazilian snail hosts of S. mansoni from available sequences of internal transcribed spacer 2 (ITS2) of the ribosomal RNA gene. Jannotti-Passos et al (2006) developed species-specific primers directed to Brazilian Biomphalaria spp.…”

Section: Introductionmentioning

confidence: 99%

Developing species-specific primers to identify Bulinus truncatus and Bulinus beccari, the intermediate hosts of Schistosoma haematobium in Saudi Arabia

Mostafa

Dajem

Al‐Qahtani

et al. 2012

Gene

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“…In order to overcome these limitations, several alternative methods for the xenomonitoring of human schistosomes have been described. Molecular approaches, such as conventional polymerase chain reaction (PCR) (14)(15)(16)(17), low-stringency polymerase chain reaction (LS-PCR) (18), restriction fragment length polymorphism (PCR-RFLP) (19), nested PCR (17), multiplex PCR (20)(21)(22), real-time quantitative PCR (qPCR) (23,24), DNA sequencing (25), and loop-mediated isothermal ampli cation (LAMP) (26)(27)(28)(29)(30) have all proved to be more accurate and sensitive alternatives than the traditional microscope-based methods. Despite the high sensitivity and speci city of molecular methods, their cost and requirement of laboratory infrastructure have limited their usage in surveillance.…”

Section: Introductionmentioning

confidence: 99%

Optimization of a Loop-Mediated Isothermal Amplification (LAMP) Assay for Schistosoma Mansoni (Trematoda: Digenea) Detection in Biomphalaria spp. from Endemic Areas for Schistosomiasis in Brazil

Mesquita

Neves

Scholte

et al. 2021

Preprint

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Background: Schistosomiasis mansoni is a neglected tropical disease endemic in Brazil caused by Schistosoma mansoni, which is transmitted by Biomphalaria snails. Among all measures to control and eliminate the disease, accurate mapping and monitoring of snail breeding sites for susceptible and/or infected hosts in endemic areas are recommended. Parasitological methods are frequently used to identify infected snails, although they have many limitations, often providing false-negative results. Loop-mediated isothermal amplification (LAMP) is a promising alternative method for a more sensitive, rapid, and cost-effective diagnosis. However, standardization of LAMP assays is challenging due to the variety of parasites that are co-endemic with S. mansoni, and their varying prevalence rates in different areas. In this work, we aimed to optimize a LAMP assay for the detection of S. mansoni in Biomphalaria snails from endemic areas in the state of Minas Gerais, Brazil. Methods: A total of 1,001 snails were collected in five municipalities of the Mucuri and Jequitinhonha Valleys. Snails were pooled and squeezed according to the collection site to detect the presence of the larval forms of S. mansoni and other trematodes. Pooled snails were submitted to pepsin digestion and DNA extraction. Then molecular assays were performed for the species-specific identification and characterization of the samples. A LAMP assay was optimized and validated using laboratory and field samples. Results: Using the parasitological method, S. mansoni cercariae were detected in snails from two collection sites. Biomphalaria glabrata, the main snail host of S. mansoni in Brazil, was detected in 72.2% of the collection sites by PCR-RFLP. Multiplex PCR, LS-PCR, and conventional PCR allowed the detection of positive snails in four additional sites. The optimized LAMP assay was effective in detecting the presence of S. mansoni infection with 100% sensitivity, 91.66% specificity, and a Kappa index of 0.88, when compared to LS-PCR and conventional PCR. Conclusions: Our findings suggest that LAMP is a good alternative for the detection and monitoring of transmission foci of S. mansoni, as it enabled the detection of infection three times more than the parasitological examination and is more applicable directly in the field when compared to other molecular approaches.

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A Multiplex-PCR approach to identification of the Brazilian intermediate hosts of Schistosoma mansoni

Abstract: Due to difficulties concerning morphological identification of planorbid snails of the genus

Cited by 9 publications

References 14 publications

Aplysina (Porifera: Demospongiae) species identification through SSCP-ITS patterns

Aplysina (Porifera: Demospongiae) species identification through SSCP-ITS patterns

Developing species-specific primers to identify Bulinus truncatus and Bulinus beccari, the intermediate hosts of Schistosoma haematobium in Saudi Arabia

Optimization of a Loop-Mediated Isothermal Amplification (LAMP) Assay for Schistosoma Mansoni (Trematoda: Digenea) Detection in Biomphalaria spp. from Endemic Areas for Schistosomiasis in Brazil

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