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Journal of Clinical Microbiology, January 2000, p. 236-240, Vol. 38, No. 1
0095-1137/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Optimized PCR Using Patient Blood Samples For Diagnosis and Follow-Up of Visceral Leishmaniasis, with Special Reference to AIDS Patients

Laurence Lachaud,1 Jacques Dereure,1 Elisabeth Chabbert,1 Jacques Reynes,2 Jean-Marc Mauboussin,3 Eric Oziol,4 Jean-Pierre Dedet,1 and Patrick Bastien1,*

Laboratoire de Parasitologie-Mycologie et Centre National de Référence sur les Leishmanioses,1 Service des Maladies Infectieuses et Tropicales,2 and Service de Médecine Interne A,4 Centre Hospitalier-Universitaire, 34000 Montpellier, and Service de Pneumologie-Médecine Interne A, Centre Hospitalier-Universitaire, 30000 Nîmes,3 France

Received 17 May 1999/Returned for modification 21 July 1999/Accepted 22 September 1999

We developed a highly sensitive PCR method that enables the diagnosis and posttherapeutic follow-up of visceral leishmaniasis with patient blood. The PCR assay was thoroughly optimized by successive procedural refinements to increase its sensitivity and specificity. It was compared to in vitro cultivation as well as to direct examination of bone marrow and to serology. Two hundred thirty-seven patients presenting with clinical signs compatible with visceral leishmaniasis were included in the study. Thirty-six were diagnosed as having Mediterranean visceral leishmaniasis (MVL). Twenty-three of them, including 19 AIDS patients, were monitored during and after treatment over a period from 2 weeks to 3 years. Our PCR assay proved more sensitive than in vitro cultivation, direct examination, and serology for all patients. It is simple and can be adapted to routine hospital diagnostic procedures. For the primary diagnosis of MVL, the sensitivity of PCR versus that of cultivation was 97 versus 55% with peripheral blood and 100 versus 81% with bone marrow samples. Regarding posttherapeutic follow-up, overall, 48% of positive samples were detected by PCR only. Seven patients presented with a clinical relapse during the study; six relapses were detected at first by PCR only, sometimes a few weeks before the reappearance of signs or symptoms. We conclude that an optimized and well-mastered PCR assay with a peripheral blood sample is sufficient to provide a secure diagnosis for all immunocompromised patients and most immunocompetent patients. We also suggest systematic posttherapeutic monitoring by PCR with peripheral blood for immunocompromised patients.

* Corresponding author. Mailing address: Laboratoire de Parasitologie-Mycologie, Centre Hospitalier-Universitaire, 163 Rue A. Broussonet, 34090 Montpellier, France. Phone: 33-4-67-63-27-51. Fax: 33-4-67-63-00-49. E-mail: genpara{at}sc.univ-montp1.fr.

Journal of Clinical Microbiology, January 2000, p. 236-240, Vol. 38, No. 1
0095-1137/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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