Rapid and simple identification of human pathogenic heterophyid intestinal fluke metacercariae by PCR-RFLP

Thaenkham, Urusa; Phuphisut, Orawan; Pakdee, Wallop; Homsuwan, Nirundorn; Sanguankiat, Surapol; Waikagul, Jitra; Nawa, Yukifumi; Dung, Do Trung

doi:10.1016/j.parint.2011.09.004

Cited by 16 publications

(7 citation statements)

References 20 publications

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“…Thus we could infer that the classification of different microalgae could effectively be done through restriction fragment length polymorphism (RFLP) analysis with the extinction enzymes Hha I and Hae III; and the taxonomic position can be accurate to Genus. The results show that the method of RFLP is simple and relatively cheap in species identification (Thaenkham et al, 2011).…”

Section: Validation Of Rflp Methodsmentioning

confidence: 92%

The biodiversity of oleaginous microalgae in Northern Qinghai-Tibet Plateau

Liu¹,

Xu²,

Yu³

et al. 2014

Afr. J. Microbiol. Res.

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Microalgae represent an exceptionally diverse but highly specialized group of micro-organisms that adapt to various ecological habitats. Due to their unique environment (extremely low temperature and anoxia), the Tibetan Plateau is among the regions with numerous rare ecotypes such as arid desert, salt marsh, alpine permafrost, hot spring and lawn. In this study, 52 microalgae from different environments in Qinghai-Tibet Plateau were isolated and divided into 11 different groups based on the results of restriction fragment length polymorphism (RFLP). Then, 2-3 strains of each group were selected for 18s rDNA molecular identification. The oil contents among the studied strains varied from 3.26 to 25.47%. The results indicate that the number and high lipid content of the strains isolated from the water were much more than those from the soil. This provides a direction for the isolation of high oleaginous microalgae.

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Section: Validation Of Rflp Methodsmentioning

confidence: 92%

The biodiversity of oleaginous microalgae in Northern Qinghai-Tibet Plateau

Liu¹,

Xu²,

Yu³

et al. 2014

Afr. J. Microbiol. Res.

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“…Many molecular techniques have been developed for the detection of a range of FBT, i.e., multiplex PCR and real-time PCR for identification of C. sinensis and O. viverrini [27,28], random amplified polymorphic DNA-PCR method for differential detection of O. viverrini and H. taichui [29], PCR targeting ITS regions for O. viverrini , C. sinensis , H. pumilio and H. taichui [30], PCR-restriction fragment length polymorphism (RFLP) for detection of O. viverrini , C. sinensis , and H. taichui [31] and for H. taichui , H. pumilio , H. yokogawai , Procerovum varium , S. falcatus , and Centrocestus formosanus [32]. Our study could identify life cycle stages of O. viverrini , eggs in infected feces, metacercariae from infected fishes, and cercarial stages from infected snails; C. sinensis eggs in infected feces as well as H. pumilio , H. taichui and S. falcatus metacercariae from infected fishes at the species level by a pyrosequencing approach.…”

Section: Discussionmentioning

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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“…Within fish-borne trematodes, Centrocestus formosanus is a small heterophyid fluke, described for the first time in Taiwan [9] and widely distributed in Asia [4,5,[10][11][12][13][14][15]. Since the 1950s, several authors have reported this species in new continents, although its occurrence could still be underestimated [16][17][18][19][20][21][22].…”

Section: Introductionmentioning

confidence: 99%

“…Subsequently, free-swimming cercariae encyst in second intermediate fish hosts, specifically in the gills, where they develop into metacercariae. Piscivorous birds and mammals, ingesting the infected fish, complete the cycle [4,15,16,18,19,23]. Analogously, human infections can occur through consumption of raw or improperly cooked fish, containing metacercariae [12,15,24].…”

Section: Introductionmentioning

confidence: 99%

“…Piscivorous birds and mammals, ingesting the infected fish, complete the cycle [4,15,16,18,19,23]. Analogously, human infections can occur through consumption of raw or improperly cooked fish, containing metacercariae [12,15,24]. C. formosanus is a generalist parasite with a broad range of fish hosts, in which it causes severe gill lesions, respiratory disorders, loss of production, and death, giving rise to health and economic concerns in relation to wild, farmed (food and ornamental), and laboratory fish [9,[17][18][19][25][26][27][28][29][30][31].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Diagnosis of Centrocestus formosanus Infection in Zebrafish (Danio rerio) in Italy: A Window to a New Globalization-Derived Invasive Microorganism

Pace

Dipineto

Aceto

et al. 2020

Animals

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Centrocestus formosanus is a digenetic trematode with a complex life cycle, involving invertebrate and vertebrate hosts, humans included. In particular, it causes gill lesions and mortality in freshwater fish species, and gastrointestinal symptoms in infected humans. Here, we describe the occurrence of C. formosanus infection in zebrafish imported in Italy and propose a newly designed species-specific primer pair to ameliorate the diagnostic investigations for C. formosanus. Gill arches of 30 zebrafish were examined for the presence of encysted metacercariae under a stereomicroscope and processed through molecular analyses targeting the ribosomal internal transcribed sequence 2 (ITS2). Although C. formosanus distribution was originally restricted to Asia, it has been subsequently reported in new countries, revealing itself as an invasive species and raising important concerns for biodiversity, economy, scientific research, as well as animal and public health. Given the crucial role played by the ornamental fish industry in spreading this parasite, there is an urgent need for control measures to prevent the introduction and establishment of C. formosanus in non-endemic areas, including Europe. We also suggest developing new strategies in microbiology and epidemiology to better explore this new globalization-derived invasive species.

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Rapid and simple identification of human pathogenic heterophyid intestinal fluke metacercariae by PCR-RFLP

Cited by 16 publications

References 20 publications

The biodiversity of oleaginous microalgae in Northern Qinghai-Tibet Plateau

The biodiversity of oleaginous microalgae in Northern Qinghai-Tibet Plateau

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

Diagnosis of Centrocestus formosanus Infection in Zebrafish (Danio rerio) in Italy: A Window to a New Globalization-Derived Invasive Microorganism

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