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Journal of Clinical Microbiology, February 2000, p. 748-751, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Use of PCR To Detect Leishmania (Viannia) spp. in Dog Blood and Bone Marrow

Richard Reithinger,1,2,* Bronwen E. Lambson,2 Douglas C. Barker,2 and Clive R. Davies1

Disease Control & Vector Biology, Department of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine, GB-London WC1E 7HT,1 and Molteno Institute for Parasitology, Department of Pathology, Cambridge University, GB-Cambridge CB2 1QP, United Kingdom2

Received 13 July 1999/Returned for modification 14 September 1999/Accepted 6 October 1999

A PCR-based protocol for the detection of Leishmania (Viannia) parasites in canine blood, buffy coat, and bone marrow was developed and was then tested with field samples taken from a random sample of 545 dogs from villages in Peru where Leishmania (Viannia) braziliensis and Leishmania (Viannia) peruviana are endemic. Comparative tests with cultured parasites mixed with dog blood showed that the PCR assay's sensitivity was significantly dependent on the DNA extraction protocol and the PCR primers used. Mass screening of field samples by the preferred PCR protocol detected American cutaneous leishmaniasis (ACL) in 44 of 545 (8.1%) dogs; 31 of 402 (7.7%), 20 of 223 (9.0%), and 8 of 46 (17.4%) were PCR positive when whole blood, buffy coat, and bone marrow aspirates, respectively, were tested. The high prevalence of Leishmania in both asymptomatic (7.6%) and symptomatic (18.0%) dogs provides further circumstantial evidence for their suspected role as reservoir hosts of ACL and indicates that hematogenous dissemination of parasites may be a more common pathological phenomenon than has previously been acknowledged. However, unlike for zoonotic visceral leishmaniasis, the comparatively low prevalence of Leishmania (Viannia) in the blood of symptomatic dogs indicates that PCR with blood cannot be the "gold standard" for the (mass) screening of samples in epidemiological studies.

* Corresponding author. Mailing address: Disease Control & Vector Biology, Department of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine, Keppel Street, GB-London WC1E 7HT, United Kingdom. Phone: 44 171 927 2350. Fax: 44 171 636 8739. E-mail: rreithinger{at}hotmail.com.

Journal of Clinical Microbiology, February 2000, p. 748-751, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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