Molecular differentiation of Trichinella spiralis, T. pseudospiralis, T. papuae and T. zimbabwensis by pyrosequencing

Sadaow, Lakkhana; Tantrawatpan, Chairat; Intapan, Pewpan M.; Lulitanond, Viraphong; Boonmars, Thidarut; Morakote, Nimit; Pozio, Edoardo; Maleewong, Wanchai

doi:10.1017/s0022149x13000308

Cited by 8 publications

(4 citation statements)

References 20 publications

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“…A polymerase chain reaction-based on the mitochondrial large subunit ribosomal RNA gene was paired with a pyrosequencing technique to distinguish the four Trichinella species, this was successfully found to be sensitive enough to identify the individual larvae [77]. PCR based on the ITS1, ITS2 and ESV regions has also been utilised for the molecular identification of the species among wildlife in South Africa [78].…”

Section: Species Identification Methodsmentioning

confidence: 99%

Non-Encapsulated Trichinella Species: T. pseudo spiralis, T. papuae and T. zimbawensis

Sharma¹,

Kaur²,

Sehgal³

2023

Cytotoxicity - Understanding Cellular Damage and Response

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Trichinellosis is a meat-borne zoonotic disease caused by nematode worms of the genus Trichinella in humans. Sylvatic animals are the main reservoir hosts of this helminth but domesticated animals, mainly swine, can also acquire the infection when they are fed with scraps of game meat. The genus used to have only one species; however, it has subsequently evolved into a multispecies genus. Due to its broad host range, it has been able to establish itself in both domestic and sylvatic cycles, allowing it to maintain a vast host reservoir. Infection has been documented in a variety of experimental species, showing that it could potentially happen in natural settings as well. Due to the considerable genetic differences among the isolates, researchers predict that the number of species and genotypes discovered within Trichinella will increase. Outbreaks caused by various species in different parts of the world have also been reported therefore prevention and control are critical in order to limit the parasite’s transmission to humans. Although molecular methods are used to identify the Trichinella species but these methods are not appropriate for the diagnosis of the infection in animals.

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Section: Species Identification Methodsmentioning

confidence: 99%

Non-Encapsulated Trichinella Species: T. pseudo spiralis, T. papuae and T. zimbawensis

Sharma¹,

Kaur²,

Sehgal³

2023

Cytotoxicity - Understanding Cellular Damage and Response

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“…Recently, DNA pyrosequencing of the PCR amplicons, the direct sequencing by the synthesis of short nucleotide fragments has been successfully applied in a variety of cases, including genotyping and species level identification of protozoan parasites [18-21], trematodes [22] and nematodes [23]. …”

Section: Introductionmentioning

confidence: 99%

Development of a PCR assay and pyrosequencing for identification of important human fish-borne trematodes and its potential use for detection in fecal specimens

et al. 2014

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BackgroundSmall liver and minute intestinal flukes are highly prevalent in Southeast Asia. Definitive diagnosis of parasite infection is usually achieved parasitologically by finding the fluke eggs in feces. However, their eggs are difficult to differentiate morphologically in fecal samples, even for experienced technicians. The present study developed a PCR assay coupled with DNA pyrosequencing for identification of the fish-borne trematodes (FBT), Opisthorchis viverrini, Clonorchis sinensis, Haplorchis taichui, H. pumilio and Stellantchasmus falcatus, and to evaluate potential detection in fecal specimens, and identification and differentiation of cercarial and metacercarial stages.MethodsPrimers targeting the partial 28S large subunit ribosomal RNA gene were designed and about 46–47 nucleotides were selected as the target region for species identification by a PCR assay coupled with a pyrosequencing technique.ResultsThe nucleotide variations at 24 positions, which is sufficient for the identification of the five species of FBT were selected. The method could identify O. viverrini and C. sinensis eggs in feces, cercarial and metacercarial stages of O. viverrini, and metacercarial stage of H. pumilio and H. taichui. The detection limit was as little as a single O. viverrini or C. sinensis egg artificially inoculated in 100 mg of non-infected fecal sample (equivalent to 10 eggs per gram), indicating highly sensitivity. The method was found to be superior to the traditional microscopy method and was more rapid than Sanger DNA sequencing.ConclusionsDNA pyrosequencing-based identification is a valuable tool for differentiating O. viverrini and other Opisthorchis-like eggs, and can be applied to epidemiological studies and for molecular taxonomic investigation of FBT in endemic areas.

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“…Pyrosequencing has been used in a broad range of applications, such as SNP genotyping, de novo mutation detection, gene identification and microbial genotyping [20]. It has recently been developed for the species-level identification of Entamoeba species [21] and has been applied to several parasite taxa, such as Plasmodium [22], Paragonimus [23] and Trichinella species [24]. The use of pyrosequencing for the identification of human taeniids has not yet been reported.…”

Section: Introductionmentioning

confidence: 99%

Rapid Molecular Identification of Human Taeniid Cestodes by Pyrosequencing Approach

et al. 2014

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Taenia saginata, T. solium, and T. asiatica are causative agents of taeniasis in humans. The difficulty of morphological identification of human taeniids can lead to misdiagnosis or confusion. To overcome this problem, several molecular methods have been developed, but use of these tends to be time-consuming. Here, a rapid and high-throughput pyrosequencing approach was developed for the identification of three human taeniids originating from various countries. Primers targeting the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of the three Taenia species were designed. Variations in a 26-nucleotide target region were used for identification. The reproducibility and accuracy of the pyrosequencing technology was confirmed by Sanger sequencing. This technique will be a valuable tool to distinguish between sympatric human taeniids that occur in Thailand, Asia and Pacific countries. This method could potentially be used for the molecular identification of the taeniid species that might be associated with suspicious cysts and lesions, or cyst residues in humans or livestock at the slaughterhouse.

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Molecular differentiation of Trichinella spiralis, T. pseudospiralis, T. papuae and T. zimbabwensis by pyrosequencing

Cited by 8 publications

References 20 publications

Non-Encapsulated Trichinella Species: T. pseudo spiralis, T. papuae and T. zimbawensis

Non-Encapsulated Trichinella Species: T. pseudo spiralis, T. papuae and T. zimbawensis

Development of a PCR assay and pyrosequencing for identification of important human fish-borne trematodes and its potential use for detection in fecal specimens

Rapid Molecular Identification of Human Taeniid Cestodes by Pyrosequencing Approach

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