Anisakis pegreffii: A quantitative fluorescence PCR assay for detection in situ

Fang, Wenzhen; Fan, Liu; Zhang, Shaolei; Lin, Jie; Xu, Shisan; Luo, Damin

doi:10.1016/j.exppara.2010.11.008

Cited by 18 publications

(13 citation statements)

References 23 publications

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“…Although several attempts to develop primers/probes to be used in RT-PCR methodology for the identification and detection of anisakids were proposed (Espiñeira et al, 2010;Mossali et al, 2010;Fang et al, 2011;Herrero et al, 2011), a species-specific RT-PCR protocol for the simultaneous detection of the most important zoonotic species present in fish of commercial value in European waters has hitherto been lacking. In addition, most of the attempts to design a protocol based on RT-PCR have been based on the ITS region of rDNA, for instance on the two Anisakis species A. simplex (s. s.) and A. pegreffii.…”

Section: Discussionmentioning

confidence: 99%

“…Similarly, the ITS region of the rDNA was used to detect anisakids in the seafood products (Espiñeira et al, 2010), but the method was not able to identify the larvae at their species level. Furthermore, a real-time PCR method for the detection in situ was proposed to amplify a region of the ITS-2 rDNA only for the species Anisakis pegreffii (Fang et al, 2011). A further probe based on the mtDNA cox1 was also proposed (Herrero et al, 2011); however, it was not able to distinguishing different species of anisakid nematodes.…”

Section: Introductionmentioning

confidence: 99%

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Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis , Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)

et al. 2018

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Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in "seafood" products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.), P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum, a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency (E) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis , Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)

et al. 2018

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“…the ISI Web of Knowledge online database was searched according to a set of search terms ( Anisakis , Anisakid) to assess the novel research articles in the field that have been published since 2011 and extract specific locality records. In total, 101 publications were considered in the present model916171822364748495051525354555657585960616263646566676869707172737475767778798081828384858687888990919293949596979899100101102103104105106107108109110111112113114115116117118119120121122123124125126127128129130131132133134135…”

Section: Methodsmentioning

confidence: 99%

Environmental variables and definitive host distribution: a habitat suitability modelling for endohelminth parasites in the marine realm

Kühn

Cunze

Kochmann

et al. 2016

Sci Rep

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Marine nematodes of the genus Anisakis are common parasites of a wide range of aquatic organisms. Public interest is primarily based on their importance as zoonotic agents of the human Anisakiasis, a severe infection of the gastro-intestinal tract as result of consuming live larvae in insufficiently cooked fish dishes. The diverse nature of external impacts unequally influencing larval and adult stages of marine endohelminth parasites requires the consideration of both abiotic and biotic factors. Whereas abiotic factors are generally more relevant for early life stages and might also be linked to intermediate hosts, definitive hosts are indispensable for a parasite’s reproduction. In order to better understand the uneven occurrence of parasites in fish species, we here use the maximum entropy approach (Maxent) to model the habitat suitability for nine Anisakis species accounting for abiotic parameters as well as biotic data (definitive hosts). The modelled habitat suitability reflects the observed distribution quite well for all Anisakis species, however, in some cases, habitat suitability exceeded the known geographical distribution, suggesting a wider distribution than presently recorded. We suggest that integrative modelling combining abiotic and biotic parameters is a valid approach for habitat suitability assessments of Anisakis, and potentially other marine parasite species.

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“…Rti-PCR methods have been reported specifically for the detection of seafoodborne nematodes (Arizono, Yamada, Tegoshi, & Yoshikawa, 2012;Fang et al, 2011). Additionally, there are Rti-PCR methods available for Giardia (Guy, Payment, Krull, & Horgen, 2003) that could be adapted to seafood matrices, if needed.…”

Section: Real-time Pcrmentioning

confidence: 99%

High throughput screening of seafood for foodborne pathogens

Jones

2015

High Throughput Screening for Food Safety Assessment

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Anisakis pegreffii: A quantitative fluorescence PCR assay for detection in situ

Cited by 18 publications

References 23 publications

Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis , Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)

Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis , Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)

Environmental variables and definitive host distribution: a habitat suitability modelling for endohelminth parasites in the marine realm

High throughput screening of seafood for foodborne pathogens

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