Sequence analysis of the β-giardin gene and development of a polymerase chain reaction–restriction fragment length polymorphism assay to genotype Giardia duodenalis cysts from human faecal samples

Cacciò, Simone M.; Giacomo, M. De; Pozio, Edoardo

doi:10.1016/s0020-7519(02)00068-1

Cited by 356 publications

(273 citation statements)

References 28 publications

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“…These were similar, but not identical, to sequences described as A2 (GenBank accession number: AY072723; Caccio et al, 2002) and A3 (GenBank accession number: AY072724; Caccio et al, 2002) but had between two and 14 single nucleotide polymorphisms (SNPs) from either A2 or A3 sequences, and all also had one SNP (at position 561) from both A2 and A3 (Table 4).…”

Section: Genotyping/sequence Analysismentioning

confidence: 70%

“…The first restriction pattern (74 bp, 126 bp, 150 bp, 201 bp, 202 bp) has been previously associated with G. duodenalis assemblage A (Caccio et al, 2002). The second pattern is similar but misses a cleavage between the fragments of 150 bp and 201 bp, thus these fragments are replaced by a single one of 351 bp.…”

Section: Rflpmentioning

confidence: 78%

“…For the remaining 16 SNP sites that could be compared, the sequence from the white-tailed deer only differed from Group BG-cer2 at nucleotide position 423, where a cytosine residue was reported. Additionally, the white-tailed deer sequence demonstrated a single SNP at nucleotide position 128, which did not occur with any of the samples isolated from G. duodenalis from moose or reindeer in any group and also does not occur in either of the documented sequences A2 and A3 (GenBank accession numbers: AY072723 and AY072724; Caccio et al, 2002).…”

Section: Discussionmentioning

confidence: 95%

“…Two genes were used for molecular characterization: the b-giardin gene and the glutamate dehydrogenase (gdh) gene, using published methods and primers (Caccio et al, 2002;Read et al, 2004), with slight modifications to described protocols. Primers, PCR reaction mixture, and reaction conditions are described in Table 2.…”

Section: Dna Isolationmentioning

confidence: 99%

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Giardia Duodenalis Cysts Isolated From Wild Moose and Reindeer in Norway: Genetic Characterization by PCR-RFLP and Sequence Analysis at Two Genes

Robertson

Forberg

Hermansen

et al. 2007

Journal of Wildlife Diseases

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ABSTRACT:There are few genotyping studies of Giardia duodenalis isolates from cervid hosts, although a previous study suggested that cervids may be a source of infection for humans and cattle. Giardia duodenalis isolates collected from wild moose (Alces alces) and reindeer (Rangifer tarandus) in Norway during 2002 and 2003 were characterized by polymerase chain reactionrestriction fraction length polymorphism (PCR-RFLP) at the b-giardin gene, and sequence analysis at both the b-giardin and glutamate dehydrogenase (gdh) genes. All results suggested that these isolates (n525) belonged to assemblage A. Three different restriction patterns were obtained with PCR-RFLP, one of which has previously been associated with assemblage A. At the b-giardin gene, sequences from six reindeer isolates and one moose isolate were identical to a previously published assemblage A sequence from G. duodenalis cysts isolated from dairy calves. The other 10 moose isolates could be divided into five groups, with between two and 14 single nucleotide polymorphisms (SNPs) from the published genotype A2. At the gdh gene, three different sequences were obtained, differing from each other by between one and 15 SNPs and which have all been previously published as genotype A1, but with different specific hosts. Grouping of the isolates based on the sequences from both genes gave complex results; whereas all the G. duodenalis isolates from reindeer grouped together, two moose isolates, which had identical sequences at the b-giardin gene, had sequences that differed from each other by 15 SNPs at the gdh gene. The results of these studies, together with the large Norwegian populations of these cervids and the amount of fecal matter they produce, indicate that moose and reindeer may be significant reservoirs of G. duodenalis infection in Norway, which may be of importance to veterinary and public health.

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Section: Genotyping/sequence Analysismentioning

confidence: 70%

Section: Rflpmentioning

confidence: 78%

Section: Discussionmentioning

confidence: 95%

Section: Dna Isolationmentioning

confidence: 99%

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Giardia Duodenalis Cysts Isolated From Wild Moose and Reindeer in Norway: Genetic Characterization by PCR-RFLP and Sequence Analysis at Two Genes

Robertson

Forberg

Hermansen

et al. 2007

Journal of Wildlife Diseases

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“…The extracted DNA was stored at -20 °C until PCR assay. The semi-nested PCR described by Caccio et al [17] with slight modifications was performed. Each reaction mixture (50 μl) contained 10 pmol of each primer (G7 and G759 or G376 and G759), 0.2 mM of each dNTPs (Fermentas, Vilnius, Lithuania), 50 mM of KCl, 10 mM of Tris-HCl (pH 9.0), 2 mM of MgCl 2 , 1.5 U of Taq DNA polymerase (Qiagen GmbH, Hilden, Germany) and 1-2 µl of DNA.…”

Section: Farm Animalsmentioning

confidence: 99%

Epidemiological survey in Łęczyńsko-Włodawskie Lake District of eastern Poland reveals new evidence of zoonotic potential of <i>Giardia intestinalis</i>

Stojecki¹,

Sroka²,

Cencek³

et al. 2015

Ann Agric Environ Med.

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Faecal samples from 297 farm animals were collected from 18 households in distinct sites of the Łęczyńsko-Włodawskie Lake District of eastern Poland. They included samples from 86 cattle (Bos taurus), 84 pigs (Sus scrofa f. domestica), 81 sheep (Ovis aries), 10 horses (Equus caballus), and 36 dogs (Canis lupus familiaris). The samples were examined for the presence of Giardia intestinalis by the Direct Fluorescence Assay (DFA) and semi-nested PCR. All amplicons were sequenced on both strands. By DFA, cysts of Giardia spp. were detected in 66 of 297 faecal samples (22.2%). Positive specimens for Giardia spp. were derived from 29.8% of examined pigs, 21.0% of sheep, 18.6% of cattle, 10% of horses, and 19.4% of dogs. Based on the detection of the β-giardin gene by PCR, 39 (13.1%) of the 297 examined samples were recognized as positive. Detection of the presence of Giardia cysts by DFA test was overall significantly higher compared to PCR (p=0.0045). By PCR, Giardia was found in 28.1% of sheep, 11.6% of cattle, 10% of horses, 9.5% of pigs and 5.6% of dogs. Partial β-giardin gene sequences were obtained for 73.7% of the PCR positive samples. From sequenced samples derived from the studied animals, Giardia were identified as assemblage A (8 samples), B (1 sample) and E (18 samples). As assemblages A and B may be zoonotic, the farm animals living in eastern Poland could be regarded as a potential source of Giardia infection for humans.

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Does urbanization ameliorate the effect of endoparasite infection in kangaroo rats?

Hurtado

Mayer

Mabry

2021

Ecology and Evolution

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Sequence analysis of the β-giardin gene and development of a polymerase chain reaction–restriction fragment length polymorphism assay to genotype Giardia duodenalis cysts from human faecal samples

Cited by 356 publications

References 28 publications

Giardia Duodenalis Cysts Isolated From Wild Moose and Reindeer in Norway: Genetic Characterization by PCR-RFLP and Sequence Analysis at Two Genes

Giardia Duodenalis Cysts Isolated From Wild Moose and Reindeer in Norway: Genetic Characterization by PCR-RFLP and Sequence Analysis at Two Genes

Epidemiological survey in Łęczyńsko-Włodawskie Lake District of eastern Poland reveals new evidence of zoonotic potential of <i>Giardia intestinalis</i>

Does urbanization ameliorate the effect of endoparasite infection in kangaroo rats?

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