This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Mathis, A. Articles by Deplazes, P. Search for Related Content PubMed PubMed Citation Articles by Mathis, A. Articles by Deplazes, P.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, 05 1995, 1145-1149, Vol 33, No. 5
Copyright © 1995 by the American Society for Microbiology. All rights reserved.

PCR and in vitro cultivation for detection of Leishmania spp. in diagnostic samples from humans and dogs

A Mathis and P Deplazes
Institute of Parasitology, University of Zurich, Switzerland.

A PCR assay for the diagnosis of leishmaniosis was developed by using primers that were selected from the sequence of the small-subunit rRNA gene. The assay was optimized for routine diagnostic use. Processing of the clinical samples is rapid and simple (lysis of erythrocytes in Tris- EDTA buffer, digestion with proteinase K directly in PCR buffer, and no further purification steps). Furthermore, an internal control is included in every specimen in order to detect the presence of PCR inhibitors. The PCR was compared with diagnostic in vitro cultivation of promastigote stages for the detection of Leishmania spp. in clinical specimens from humans and dogs with a tentative diagnosis of leishmaniosis. PCR and cultivation gave identical results with all but 1 of the 95 specimens from humans. The PCR result in this case was false negative, possibly because of unequal apportionment of this sample. With 10 skin biopsies from six patients with cutaneous leishmaniosis, the sensitivity was 60%. For six human immunodeficiency virus-positive patients with visceral leishmaniosis, all bone marrow biopsies and 7 of 11 whole blood samples (after isolation of leukocytes by Ficoll-Paque) were positive in both tests. PCR detected one more case with the use of 500 microliters of whole blood with direct lysis of the erythrocytes in Tris-EDTA buffer. With dog lymph node aspirates, the sensitivity was 100% (16 of 16 samples) for both methods; furthermore, PCR was positive for 5 of 13 whole blood samples from dogs with leishmaniosis. The specificity of the PCR was 100% (70 specimens from patients without leishmaniosis).(ABSTRACT TRUNCATED AT 250 WORDS)

This article has been cited by other articles:

• Alvar, J., Aparicio, P., Aseffa, A., Den Boer, M., Canavate, C., Dedet, J.-P., Gradoni, L., Ter Horst, R., Lopez-Velez, R., Moreno, J. (2008). The Relationship between Leishmaniasis and AIDS: the Second 10 Years. Clin. Microbiol. Rev. 21: 334-359 [Abstract] [Full Text]  
• Mettler, M., Grimm, F., Capelli, G., Camp, H., Deplazes, P. (2005). Evaluation of Enzyme-Linked Immunosorbent Assays, an Immunofluorescent-Antibody Test, and Two Rapid Tests (Immunochromatographic-Dipstick and Gel Tests) for Serological Diagnosis of Symptomatic and Asymptomatic Leishmania Infections in Dogs. J. Clin. Microbiol. 43: 5515-5519 [Abstract] [Full Text]  
• Herwegh, S., Carnoy, C., Wallet, F., Loiez, C., Courcol, R. J. (2005). Development and Use of an Internal Positive Control for Detection of Bordetella pertussis by PCR. J. Clin. Microbiol. 43: 2462-2464 [Abstract] [Full Text]  
• KATO, H., UEZATO, H., KATAKURA, K., CALVOPINA, M., MARCO, J. D., BARROSO, P. A., GOMEZ, E. A., MIMORI, T., KORENAGA, M., IWATA, H., NONAKA, S., HASHIGUCHI, Y. (2005). DETECTION AND IDENTIFICATION OF LEISHMANIA SPECIES WITHIN NATURALLY INFECTED SAND FLIES IN THE ANDEAN AREAS OF ECUADOR BY A POLYMERASE CHAIN REACTION. Am J Trop Med Hyg 72: 87-93 [Abstract] [Full Text]  
• Fissore, C., Delaunay, P., Ferrua, B., Rosenthal, E., Del Giudice, P., Aufeuvre, J.-P., Le Fichoux, Y., Marty, P. (2004). Convenience of Serum for Visceral Leishmaniasis Diagnosis by PCR. J. Clin. Microbiol. 42: 5332-5333 [Abstract] [Full Text]  
• Marfurt, J., Nasereddin, A., Niederwieser, I., Jaffe, C. L., Beck, H.-P., Felger, I. (2003). Identification and Differentiation of Leishmania Species in Clinical Samples by PCR Amplification of the Miniexon Sequence and Subsequent Restriction Fragment Length Polymorphism Analysis. J. Clin. Microbiol. 41: 3147-3153 [Abstract] [Full Text]  
• Gangneux, J.-P., Menotti, J., Lorenzo, F., Sarfati, C., Blanche, H., Bui, H., Pratlong, F., Garin, Y.-J.-F., Derouin, F. (2003). Prospective Value of PCR Amplification and Sequencing for Diagnosis and Typing of Old World Leishmania Infections in an Area of Nonendemicity. J. Clin. Microbiol. 41: 1419-1422 [Abstract] [Full Text]  
• Reithinger, R., Espinoza, J. C., Courtenay, O., Davies, C. R. (2003). Evaluation of PCR as a Diagnostic Mass-Screening Tool To Detect Leishmania (Viannia) spp. in Domestic Dogs (Canis familiaris). J. Clin. Microbiol. 41: 1486-1493 [Abstract] [Full Text]  
• Schulz, A., Mellenthin, K., Schonian, G., Fleischer, B., Drosten, C. (2003). Detection, Differentiation, and Quantitation of Pathogenic Leishmania Organisms by a Fluorescence Resonance Energy Transfer-Based Real-Time PCR Assay. J. Clin. Microbiol. 41: 1529-1535 [Abstract] [Full Text]  
• Rodrigues, E. H. G., Felinto de Brito, M. E., Mendonca, M. G., Werkhauser, R. P., Coutinho, E. M., Souza, W. V., Militao de Albuquerque, M. d. F. P., Jardim, M. L., Abath, F. G. C. (2002). Evaluation of PCR for Diagnosis of American Cutaneous Leishmaniasis in an Area of Endemicity in Northeastern Brazil. J. Clin. Microbiol. 40: 3572-3576 [Abstract] [Full Text]  
• Bulle, B., Millon, L., Bart, J.-M., Gallego, M., Gambarelli, F., Portus, M., Schnur, L., Jaffe, C. L., Fernandez-Barredo, S., Alunda, J. M., Piarroux, R. (2002). Practical Approach for Typing Strains of Leishmania infantum by Microsatellite Analysis. J. Clin. Microbiol. 40: 3391-3397 [Abstract] [Full Text]  
• Lachaud, L., Marchergui-Hammami, S., Chabbert, E., Dereure, J., Dedet, J. P., Bastien, P. (2002). Comparison of Six PCR Methods Using Peripheral Blood for Detection of Canine Visceral Leishmaniasis. J. Clin. Microbiol. 40: 210-215 [Abstract] [Full Text]  
• Salotra, P., Sreenivas, G., Pogue, G. P., Lee, N., Nakhasi, H. L., Ramesh, V., Negi, N. S. (2001). Development of a Species-Specific PCR Assay for Detection of Leishmania donovani in Clinical Samples from Patients with Kala-Azar and Post-Kala-Azar Dermal Leishmaniasis. J. Clin. Microbiol. 39: 849-854 [Abstract] [Full Text]  
• Lachaud, L., Chabbert, E., Dubessay, P., Reynes, J., Lamothe, J., Bastien, P. (2001). Comparison of Various Sample Preparation Methods for PCR Diagnosis of Visceral Leishmaniasis Using Peripheral Blood. J. Clin. Microbiol. 39: 613-617 [Abstract] [Full Text]  
• Lachaud, L., Dereure, J., Chabbert, E., Reynes, J., Mauboussin, J.-M., Oziol, E., Dedet, J.-P., Bastien, P. (2000). Optimized PCR Using Patient Blood Samples For Diagnosis and Follow-Up of Visceral Leishmaniasis, with Special Reference to AIDS Patients. J. Clin. Microbiol. 38: 236-240 [Abstract] [Full Text]  
• Pirmez, C., da Silva Trajano, V., Neto, M. P.-O., da-Cruz, A. M., Gonçalves-da-Costa, S. C., Catanho, M., Degrave, W., Fernandes, O. (1999). Use of PCR in Diagnosis of Human American Tegumentary Leishmaniasis in Rio de Janeiro, Brazil. J. Clin. Microbiol. 37: 1819-1823 [Abstract] [Full Text]  
• Harris, E., Kropp, G., Belli, A., Rodriguez, B., Agabian, N. (1998). Single-Step Multiplex PCR Assay for Characterization of New World Leishmania Complexes. J. Clin. Microbiol. 36: 1989-1995 [Abstract] [Full Text]  
• Tang, Y.-W., Procop, G. W., Persing, D. H. (1997). Molecular diagnostics of infectious diseases. Clin. Chem. 43: 2021-2038 [Abstract] [Full Text]