PCR-based methods for identification of potentially zoonotic ascaridoid parasites of the dog, fox and cat

Jacobs, D. Ε.; Zhu, Xingquan; Gasser, Robin B.; Chilton, Neil B.

doi:10.1016/s0001-706x(97)00093-4

Cited by 155 publications

(112 citation statements)

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“…In comparison, the second internal transcribed spacer (ITS-2) region, an insertion between the 5.8S and large subunit rRNA gene, has proven to be particularly valuable in resolving the taxonomic status of various parasitic groups including cestodes (Bowles et al 1995), trematodes (Blair et al 1996) and nematodes (Hoste et al 1995, Stevenson et al 1995. ITS-2 sequence has been utilised to identify single eggs of strongyloid nematodes to the species level (Campbell et al 1995) and for differentiating ascaridoid infections in dog, fox and cat (Jacobs et al 1997).…”

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confidence: 99%

“…ITS-2 of T. leonina, T. cati and T. canis were amplified as described by Jacobs et al (1997) using three primer sets Tleo1(5'-CGAACGCTCATATAACGGCATACTC-3')-NC2 (5'-TTAGTTTCTTTTCCTCCGCT-3'), Tcat1(5' GGAGAAG TAAGATCGTGGCACGCGT-3')-NC2, and Tcan1(5'-AG-TATGATGGGCGCGCCAAT-3')-NC2. The anticipated size of the ITS-2 amplified fragments of T. leonina, T. cati and T. canis were 300 bp, 370 bp and 380 bp, respectively (Jacobs et al 1997).…”

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confidence: 99%

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Detection and molecular characterization of ascarid nematode infection (Toxascaris leonina and Toxocara cati) in captive Asiatic lions (Panthera leo persica)

Pawar

Lakshmikantan

Hasan

et al. 2012

Acta Parasitologica

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The objective of this study was to investigate the ascarid infection in Asiatic lions using scat samples, based on microscopic analysis, PCR amplification of the ITS-2 region of ribosomal DNA and sequence analysis of the amplicons. Microscopic analysis indicated the presence of eggs of Toxascaris leonina in eleven of the sixteen scat samples analysed and in one of these eleven scats eggs of Toxocara cati were also detected. In five of the scats eggs were not detectable. The presence of T. leonina in all the infected samples was also confirmed by PCR amplification of the ITS-2 of ribosomal RNA gene and five of these also showed amplicons corresponding to T. cati, respectively. Toxocara canis infection was not observed in any of the scat samples. Nucleotide sequence analysis of the ITS-2 region indicated 97% to 99% similarity with T. leonina and T. cati, respectively. To our knowledge, this is the first molecular characterization of ascarid infection in captive Asiatic lions from a zoological garden of India. This study also indicates that Asiatic lions are more prone to infection either with T. leonina or T. cati and the parasite is not host specific.

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confidence: 99%

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confidence: 99%

Detection and molecular characterization of ascarid nematode infection (Toxascaris leonina and Toxocara cati) in captive Asiatic lions (Panthera leo persica)

Pawar

Lakshmikantan

Hasan

et al. 2012

Acta Parasitologica

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“…1). In previous investigations (Jacobs et al, 1997;Zhu et al, 1998) genomic DNA was extracted from adult worms and embryonated eggs. Our innovation was to adapt the PCR for detection of T. canis gDNA extracted from a tissue of experimentally infected animals.…”

Section: Resultsmentioning

confidence: 99%

“…The extracted DNA was used for PCR, in which amplification of internal transcribed spacer 2 (ITS-2) was performed according to Jacobs et al (1997). The region spanning ITS-2 was amplified from the gDNA (5-10ng) by PCR using oligonucleotide specific for T. canis: Tcan1 and NC2.…”

Section: Pcr Methodsmentioning

confidence: 99%

“…To support the diagnosis, detection of the parasite DNA in a patient tissue sample with molecular technique based on the polymerase chain reaction (PCR) may be involved. Methods designed to discriminate T. canis and T. cati are based on PCR analysis of genomic DNA extracted from adult worms or embryonated eggs collected from the uteri of female worms (Jacobs et al, 1997;Zhu et al, 1998). We have used a PCR-restriction fragment length polymorphism (RFLP) technique for the differentiation of T. canis and T. cati eggs isolated from soil (Borecka, 2004).…”

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confidence: 99%

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Detection of Toxocara canis larvae by PCR in the liver of experimentally infected Mongolian gerbils (Meriones unguiculatus)

et al. 2008

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SummaryToxocariasis is a common human zoonosis, which induces a clinically unapparent course of infection. Diagnosis is difficult and relies upon serological testing (searching of specific IgG antibodies by ELISA), laboratory abnormalities and clinical manifestations. The polymerase chain reaction (PCR) technique was adapted for the detection of Toxocara canis larvae in a host tissue. Mongolian gerbils (Meriones unguiculatus) were used as an animal model for human toxocariasis. 8 animals were inoculated with 1000 T. canis eggs, four uninfected were used as control. At 3, 5, 7, and 14 days post-infection, 2 infected and 1 control gerbil were killed and their livers were used for molecular analysis. Specific primer in the PCR reaction allowed identification of T. canis larvae, with the parasite gDNA found in the liver of all infected gerbils. The results indicate that the PCR method has a potential as a supporting technique for the diagnosis of human toxocariasis.

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Mutation scanning for sequence variation in three mitochondrial DNA regions for members of theContracaecum osculatum (Nematoda: Ascaridoidea) complex

Hu¹,

D’Amelio²,

Zhu³

et al. 2001

Electrophoresis

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Anisakid nematodes of seals from different geographical origins, previously identified by multilocus enzyme electrophoresis as Contracaecum osculatum A (CoA), C. osculatum B (CoB), C. osculatum C (CoC), C. osculatum D (CoD), C. osculatum E (CoE) and C. osculatum baicalensis (Cob), were characterised genetically using a mutation scanning approach, in order to define genetic markers for their specific identification and differentiation. Three mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit I (COI), and the small and large subunits of rRNA (ssrRNA and IsrRNA, respectively) were amplified separately from individual nematodes by polymerase chain reaction (PCR), analysed by single-strand conformation polymorphism (SSCP), and samples displaying sequence variability were subjected to sequencing. Forty-six haplotypes were defined for 62-66 individuals (representing the six members of C. osculatum). All taxa except CoD and CoE could be identified, or delineated from one another, by nucleotide differences in the COI, ssrRNA and/or IsrRNA sequences. For all three mtDNA regions, 4 (10.5%), 7 (18.4%), 15 (39.5%) and 11 (28.9%) of 38 nucleotide positions were considered diagnostic (fixed) and could thus unequivocally delineate CoA, CoB, CoC and Cob. The lack of an unequivocal nucleotide difference in any of the three mtDNA sequences between CoD and CoE was in accordance with previous ribosomal DNA sequence data but inconsistent with multilocus enzyme electrophoretic data. Using all fixed nucleotide positions, CoA, CoD/E and CoB were genetically more similar to Cob than each was to CoC, similar to previous findings. In spite of not being able to distinguish among all six taxa of C. osculatum, the present study demonstrated clearly the usefulness and attributes of the mutation scanning approach for investigating population genetic structures of species of parasitic nematodes.

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PCR-based methods for identification of potentially zoonotic ascaridoid parasites of the dog, fox and cat

Cited by 155 publications

References 14 publications

Detection and molecular characterization of ascarid nematode infection (Toxascaris leonina and Toxocara cati) in captive Asiatic lions (Panthera leo persica)

Detection and molecular characterization of ascarid nematode infection (Toxascaris leonina and Toxocara cati) in captive Asiatic lions (Panthera leo persica)

Detection of Toxocara canis larvae by PCR in the liver of experimentally infected Mongolian gerbils (Meriones unguiculatus)

Mutation scanning for sequence variation in three mitochondrial DNA regions for members of theContracaecum osculatum (Nematoda: Ascaridoidea) complex

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