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Polymerase chain reaction—identification of Dirofilaria repens and Dirofilaria immitis

Published online by Cambridge University Press:  06 April 2009

G. Favia ,
A. Lanfrancotti ,
A. Della Torre ,
G. Cancrini  and
M. Coluzzi
G. Favia*
Affiliation:
Istituto di Parassitologia Fondazione Pasteur-Cenci Bolognetti, Università ‘La Sapienza’, Piazzale Aldo Moro 5, 00185 Rome, Italy
A. Lanfrancotti
Affiliation:
Istituto di Parassitologia Fondazione Pasteur-Cenci Bolognetti, Università ‘La Sapienza’, Piazzale Aldo Moro 5, 00185 Rome, Italy
A. Della Torre
Affiliation:
Istituto di Parassitologia Fondazione Pasteur-Cenci Bolognetti, Università ‘La Sapienza’, Piazzale Aldo Moro 5, 00185 Rome, Italy
G. Cancrini
Affiliation:
Istituto di Parassitologia Fondazione Pasteur-Cenci Bolognetti, Università ‘La Sapienza’, Piazzale Aldo Moro 5, 00185 Rome, Italy
M. Coluzzi
Affiliation:
Istituto di Parassitologia Fondazione Pasteur-Cenci Bolognetti, Università ‘La Sapienza’, Piazzale Aldo Moro 5, 00185 Rome, Italy
*
* Corresponding author. Tel: +39 6 49914900. Fax: + 39 6 49914644. E-mail: favia@axrma.uniromal.it
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Summary

On the basis of known DNA sequences of Dirofilaria repens and D. immitis we designed specific primers for the amplification by Polymerase Chain Reaction (PCR) of the DNA from the two species. The PCR-based identification was found to be unambiguous and allowed specific diagnosis of microfilariae in blood samples, of developing larvae in the mosquito vector and of immature adults in bioptic material, overcoming the serious constraints of the morphological separation of these filarial parasites at the pre-adult stages. The technique was found to be very sensitive and applicable to samples stored either dry or in various preservation media, with the exception of formalin. The reliable identification of D. repens and D. immitis from bioptic material is expected to greatly enhance the chances of detecting human infections and to further clarify the role of the two parasites as pathogens of man. The possibility of routine identification of developing larvae in the vector will substantially improve the perspectives for epidemiological investigations, particularly in Southern European regions, such as Italy, where the two nematode species are largely sympatric.

Keywords

Dirofilaria repensDirofilaria immitispolymerase chain reactiondiagnostic methodmosquitoes
Type
Research Article
Information
Parasitology , Volume 113 , Issue 6 , December 1996 , pp. 567 - 571
Copyright
Copyright © Cambridge University Press 1996

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References

REFERENCES

Boreham, P. F. L. (1988). Dirofilariasis in man. In Dirofilariasis (ed. Boreham, P. F. L. & Atwell, R. B.), pp. 217–226. CRC Press, Florida.Google Scholar
Cancrini, G., D'Amelio, S., Mattiucci, S. & Coluzzi, M. (1991). Identification of Dirofilaria in man by multilocus electrophoretic analysis. Annals of Tropical Medicine and Parasitology, 5, 529–532.CrossRefGoogle Scholar
Cancrini, G., Tringali, R. & Buccheri, G. (1996). Dirofilariosi umana: tre nuovi casi osservati in Sicilia e problemi diagnostici. Giornale Italiano di Malattie Infettive 2, 110–114.Google Scholar
Chandrasekharan, N. V., Karunanayake, E. H., Franzen, L., Abeyewick-Reme, W. & Petterson, U. (1994). Dirofilaria repens: cloning and characterization of a repeated DNA sequence for the diagnosis of dirofilariasis in dogs, Canis familiaris. Experimental Parasitology 78, 279–286.CrossRefGoogle ScholarPubMed
Favia, G., Dimopoulous, G., Della, Torre A., Touré, Y. T., Coluzzi, M. & Louis, C. (1994). Polymorphisms detected by random PCR distinguish between chromosomal forms of Anopheles gambiae. Proceedings of the National Academy of Sciences, USA 91, 10315–10319.CrossRefGoogle ScholarPubMed
Genchi, C., Di Sacco, B. & Cancrini, G. (1992). Epizootiology of canine and feline heartworm infections in Northern Italy: possible mosquito vectors. Proceedings of the 7th Triennial Heartzvorm Symposium 1992, 39–46.Google Scholar
Hughes, M. A. & Crampton, J. M. (1993). The development and use of repetitive sequences as DNA probes for parasite detection and species identification. Methods in Molecular Biology 21, 169–189.Google ScholarPubMed
Lock, J. B. (1988). Dirofilaria sp.: taxonomy and distribution. In Dirofilariasis (ed. Boreham, P. F. L. & Atwell, R. B.), pp. 1–28. CRC Press, Florida.Google Scholar
Nozais, J. P., Bain, O. & Gentilini, M. (1994). Un Cas de dirofilariose sous-cutanée a Dirofilaria (Nochtiella) repens avec microfilaremie en provenance de Corse. Bulletin de la Société de Pathologie Exotique 87, 183–185.Google Scholar
Pampiglione, S., Canestri, Trotti G. & Rivasi, F. (1995). Human dirofilariasis due to Dirofilaria (Nochtiella) repens: a review of world literature. Parassitologia 37, 148–191.Google ScholarPubMed
Poole, C. B., IIIGrandea, A. G., Maina, C. V., Jenkins, R. E., Selkirk, M. E. & McReynolds, L. A. (1992). Cloning of a cuticular antigen that contains multiple tandem repeats from the filarial parasite Dirofilaria immitis. Proceedings of the National Academy of Sciences, USA 89, 5986–5990.CrossRefGoogle ScholarPubMed
Scoles, G. A. & Kambhampati, S. (1995). Polymerase chain reaction-based method for the detection of canine heartworm (Filarioidea: Onchocercidae) in mosquitoes (Diptera: Culicidae) and vertebrate hosts. Journal of Medical Entomology 32, 864–869.CrossRefGoogle ScholarPubMed
Siridewa, K., Karunanayake, E. H. & Chandrasekharan, N. V. (1996). Polymerase chain reaction-based technique for the detection of Wuchereria bancrofti in human blood samples, hydrocele fluid, and mosquito vectors. American Journal of Tropical Medicine and Hygiene 54, 72–76.CrossRefGoogle ScholarPubMed
Smith, J. D. (1995). Rare, new and emerging helminth zoonoses. Advances in Parasitology 36, 19–40.Google Scholar
Sutton, R. H. (1988). Pathology and pathogenesis of Dirofilariasis. In Dirofilariasis (ed. Boreham, P. F. & Atwell, R. B.), pp. 100–132. CRC Press, Boca Raton, Florida.Google Scholar