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Multiplex PCR for differential identification of broad tapeworms (Cestoda: Diphyllobothrium) infecting humans.

ABSTRACT: The specific identification of broad tapeworms (genus Diphyllobothrium) infecting humans is very difficult to perform by morphological observation. Molecular analysis by PCR and sequencing represents the only reliable tool to date to identify these parasites to the species level. Due to the recent spread of human diphyllobothriosis in several countries, a correct diagnosis has become crucial to better understand the distribution and the life cycle of human-infecting species as well as to prevent the introduction of parasites to disease-free water systems. Nevertheless, PCR and sequencing, although highly precise, are too complicated, long, and expensive to be employed in medical laboratories for routine diagnostics. In the present study we optimized a cheap and rapid molecular test for the differential identification of the most common Diphyllobothrium species infecting humans (D. latum, D. dendriticum, D. nihonkaiense, and D. pacificum), based on a multiplex PCR with the cytochrome c oxidase subunit 1 gene of mitochondrial DNA.

SUBMITTER: Wicht B

PROVIDER: S-EPMC2937707 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature

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Multiplex PCR for differential identification of broad tapeworms (Cestoda: Diphyllobothrium) infecting humans.

Wicht Barbara B Yanagida Tetsuya T Scholz Tomás T Ito Akira A Jiménez Juan A JA Jiménez Juan A JA Brabec Jan J

Journal of clinical microbiology 20100630 9

The specific identification of broad tapeworms (genus Diphyllobothrium) infecting humans is very difficult to perform by morphological observation. Molecular analysis by PCR and sequencing represents the only reliable tool to date to identify these parasites to the species level. Due to the recent spread of human diphyllobothriosis in several countries, a correct diagnosis has become crucial to better understand the distribution and the life cycle of human-infecting species as well as to prevent ...[more]

PMID: 20592146

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Project description:BackgroundA total number of 14 valid species of Diphyllobothrium tapeworms have been described in literature to be capable of causing diphyllobothriosis, with D. latum being the major causative agent of all human infections. However, recent data indicate that some of these infections, especially when diagnosed solely on the basis of morphology, have been identified with this causative agent incorrectly, confusing other Diphyllobothrium species with D. latum. Another widely distributed species, D. dendriticum, has never been considered as a frequent parasite of man, even though it is found commonly throughout arctic and subarctic regions parasitizing piscivorous birds and mammals. Recent cases of Europeans infected with this cestode called into question the actual geographic distribution of this tapeworm, largely ignored by medical parasitologists.Methodology and resultsOn the basis of revision of more than 900 available references and a description and revision of recent European human cases using morphological and molecular (cox1) data supplemented by newly characterized D. dendriticum sequences, we updated the current knowledge of the life-cycle, geographic distribution, epidemiological status, and molecular diagnostics of this emerging causal agent of zoonotic disease of man.ConclusionsThe tapeworm D. dendriticum represents an example of a previously neglected, probably underdiagnosed parasite of man with a potential to spread globally. Recent cases of diphyllobothriosis caused by D. dendriticum in Europe (Netherlands, Switzerland and Czech Republic), where the parasite has not been reported previously, point out that causative agents of diphyllobothriosis and other zoonoses can be imported throughout the world. Molecular tools should be used for specific and reliable parasite diagnostics, and also rare or non-native species should be considered. This will considerably help improve our knowledge of the distribution and epidemiology of these human parasites.

Project description:Gyrocotylideans are evolutionary ancient parasitic flatworms, and like their hosts-a relict group of holocephalan fishes (Chimaeriformes)-they are considered to be "living fossils" of a vanished past. However, the species diversity, host associations and biogeography of these most basal tapeworms are poorly known. Herein, we provide evidence of a conspicuous contrast between the genetic and morphological data based on an examination of newly collected and properly processed Gyrocotyle specimens (hologenophores) isolated from holocephalans off Taiwan and Argentina. Our molecular data, inferred from three genes (COI, 28S rRNA, 18S rRNA), showed unexpected genetic interrelationships among isolates of the genus Gyrocotyle, because each of the four genotypes from Taiwan clustered with isolates of distinct gyrocotylideans from the North Atlantic. Three genotypes of Gyrocotyle from Taiwan were morphologically almost indistinguishable from each other but represented distinct genetic lineages; a single specimen of Gyrocotyle sp. genotype 4 exhibited a clear genetic and morphological distinctness, though its formal description as a new species would be premature. Additionally, specimens of Gyrocotyle rugosa Diesing, 1850, from the type host Callorhinchus callorynchus from Argentina, provided the first genetic data on the type species of the genus and enabled us to characterise it, which is necessary for future taxonomic studies. The finding of some specimens of Gyrocotyle sp. genotype 3 in Chimaera phantasma, and another one in C. cf. argiloba, together with the putative conspecificity of an unidentified gyrocotylidean from Callorhinchus milii off Australia and G. rugosa from C. callorynchus off Argentina, represent evidence that one gyrocotylidean species may parasitise more than one holocephalan host species. Existing taxonomic problems and conflicts between morphological and molecular data on species of Gyrocotyle can only be resolved if hologenophores from type hosts and localities of nominal taxa are properly characterised genetically and morphologically.

Dataset Information

Multiplex PCR for differential identification of broad tapeworms (Cestoda: Diphyllobothrium) infecting humans.

Publications

Multiplex PCR for differential identification of broad tapeworms (Cestoda: Diphyllobothrium) infecting humans.

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