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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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