Detection of Lymnaea columella infection by Fasciola hepatica through Multiplex-PCR

Magalhães, Kelly Grace; Passos, Liana Konovaloff Jannotti; Carvalho, Omar dos Santos

doi:10.1590/s0074-02762004000400013

Cited by 32 publications

(29 citation statements)

References 22 publications

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“…Amplification of F. hepatica specific mDNA was achieved and a ladder pattern with intervals of 85 nucleotides was visible. Such pattern mirrored the amplification of a tandem repeat DNA region (Magalhaes et al 2004). The reaction was sensitive, as it detected a single miracidium (0.8 ng/µl) of F. hepatica in the presence of snail DNA (7 ng/µl).…”

Section: Multiplex Pcrmentioning

confidence: 81%

“…Specific amplification was obtained in infected snails as soon as 1 day post-exposure. The designed primers were specific for F. hepatica detection, as demonstrated by the band profiles obtained from snails infected with this trematode and other species commonly found in P. columella (Magalhaes et al 2004). …”

Section: Multiplex Pcrmentioning

confidence: 99%

“…This technique needs as much pairs of primers as sequences to be amplified. Mitochondrial DNA (mDNA) was used in a multiplex PCR technique to identify larval stages of F. hepatica in experimentally infected snails in Brazil (Magalhaes et al 2004). From another complete mitochondrial DNA sequence of F. hepatica (Genbank accession number, AF216697), specific primers were designed for a conserved and repetitive region of this trematode.…”

Section: Multiplex Pcrmentioning

confidence: 99%

“…From another complete mitochondrial DNA sequence of F. hepatica (Genbank accession number, AF216697), specific primers were designed for a conserved and repetitive region of this trematode. A pair of primers was used in F. hepatica experimentally infected P. columella during the prepatent period simultaneously with another pair of primers which amplified, under high stringency conditions, the internal transcribed spacer (ITS) region of the trematode and the mollusc rDNA, thus acting as an internal control (Magalhaes et al 2004). PCR products were visualized on 6% silver-stained polyacrylamide gels.…”

Section: Multiplex Pcrmentioning

confidence: 99%

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The detection and quantification of a digenean infection in the snail host with special emphasis on Fasciola sp.

2008

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In this review, ten methods used to study digenean infections in their intermediate hosts were compared to determine which one should be used either in the field or in the lab to establish the prevalence and intensity of infections in snails. Snail crushing and snail dissection allow quick establishing of prevalence in natural or experimental infections, whereas histology is considered as the most accurate approach to assess the intensity of infection. The follow-up of cercarial shedding only gave an idea on cercarial production. Among recently developed techniques, polymerase chain reaction (PCR) brings the most accurate information and shows high sensitivity and specificity levels when compared to blotting techniques. The easiness and relatively low cost of the basic PCR protocol make it interesting to investigate the epidemiology of the liver fluke in a lab with limited financial resources. Nevertheless, if this technique allows a relatively good estimation of the prevalence, information concerning the intensity of infection is best obtained through real time PCR. However, at the time being this technique is too expensive to be used routinely in the field. The choice between classical or new techniques is usually based on a compromise, as each technique has its advantages and drawbacks.

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Section: Multiplex Pcrmentioning

confidence: 81%

Section: Multiplex Pcrmentioning

confidence: 99%

Section: Multiplex Pcrmentioning

confidence: 99%

Section: Multiplex Pcrmentioning

confidence: 99%

See 2 more Smart Citations

The detection and quantification of a digenean infection in the snail host with special emphasis on Fasciola sp.

2008

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“…Before cercarial development, however, the intra-molluscan stages of the different trematode species are difficult to differentiate (Kaplan et al, 1995). For this reason, several DNA-or RNA-based techniques were developed (Shubkin et al, 1992;Rognlie et al, 1994;Kaplan et al, 1995;Kramer and Schnieder, 1998;Mostafa et al, 2003;Magalhaes et al, 2004;Cucher et al, 2006). These techniques are specific and sensitive (Kaplan et al, 1997) but are rarely used to detect naturally occurring infections, although this should be one of their main applications.…”

Section: Introductionmentioning

confidence: 99%

Fasciola hepatica: An assessment on the vectorial capacity of Radix labiata and R. balthica commonly found in Belgium

Caron¹,

Lasri²,

Losson³

2007

Veterinary Parasitology

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A previous study conducted in Belgium revealed that genetic material of Fasciola sp. was present in snail species belonging to the genus Radix. Here, these snails were collected and identified by DNA-based techniques as Radix labiata and Radix balthica. These two species and Galba truncatula (the major intermediate host in Europe) were experimentally infected with Fasciola hepatica. The resulting metacercariae were fed to rats and the infection was monitored using several techniques. Microscopy revealed the presence of larval stages in 78.3, 45, and 6.25% of G. truncatula, R. labiata, and R. balthica snails, respectively. These results were confirmed by a PCR that amplifies a Fasciola sp. specific sequence. Furthermore, this PCR was found to be more sensitive than microscopic examination. R. labiata shed fewer metacercariae than G. truncatula but these were as infective to rats as those shed by G. truncatula. This study demonstrates that R. labiata may act as an incidental intermediate host for F. hepatica in Belgium. #

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A pooling strategy of a PCR-based assay to detect Angiostrongylus cantonensis in snail intermediate host, Pomacea canaliculata

Wei

Liu

et al. 2010

Front. Med. China

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Pooling field specimens could reduce the number of assay and thus increase the efficiency in detecting and screening pathogen infections by polymerase chain reaction (PCR)-based assay. We investigated a pooling strategy in diagnosis of Angiostrongylus cantonensis in Pomacea canaliculata. Two settings of specimens were prepared, divided into portions and detected by multiplex PCR. Specimens A was 0.4490 g positive lung tissue of 28 larval nodes from four snails mixed with 1.310 g negative lung tissue from six snails and divided into 32 portions. Specimens B was 0.5448 g positive lung tissue with 26 larval nodes from two snails mixed with 1.092 g negative lung tissue from seven snails and divided into 48 portions. Repeated sampling was performed and sample size-accumulated positive rate curves were drawn. According to the sample size-accumulated positive rate curves, the appropriate sample size of the two specimens were 18 and 15, respectively, which is 0.36-0.58 to the total sample size. These test characteristics and the relevant factors to the sample size would need to be determined in much larger studies and more appropriately in field populations. The result indicates that the number of larval node is not the most important, nor the only factor to the sample size. It also implies the feasibility to detect A. cantonensis in P. canaliculata by pooling strategies.

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Detection of Lymnaea columella infection by Fasciola hepatica through Multiplex-PCR

Cited by 32 publications

References 22 publications

The detection and quantification of a digenean infection in the snail host with special emphasis on Fasciola sp.

The detection and quantification of a digenean infection in the snail host with special emphasis on Fasciola sp.

Fasciola hepatica: An assessment on the vectorial capacity of Radix labiata and R. balthica commonly found in Belgium

A pooling strategy of a PCR-based assay to detect Angiostrongylus cantonensis in snail intermediate host, Pomacea canaliculata

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