Affiliation:
1. Department of Medical Parasitology and Infection Biology, Swiss Tropical Institute, Basel, Switzerland
2. Department of Parasitology, Hadassah Medical School, Hebrew University, Jerusalem, Israel
Abstract
ABSTRACT
We recently developed a new PCR-restriction fragment length polymorphism (RFLP)-based assay using the miniexon sequence from the genus
Leishmania
. Here we report the application of this new genotyping method to naturally infected clinical samples for the differentiation of New and Old World
Leishmania
species. Of the newly developed assay and four currently applied diagnostic tests (i.e., in vitro cultivation, serology, and two other molecular assays using either the small subunit-internal transcribed spacer sequence or a repetitive genomic sequence), the miniexon assay showed the highest sensitivity, 89.7%, compared to 70.6, 57.1, 51.7, and 79.3%, respectively. Species differentiation was robust and reliable compared with that by two other
Leishmania
genotyping techniques. The assay provides a valuable tool for the identification of
Leishmania
directly from clinical samples and enables determination of the infecting species by a facile technique with high discrimination power. Since
Leishmania
causes a broad spectrum of diseases distinguished by different parasite and host factors, detection and characterization of the infecting species is crucial for the confirmation of a diagnosis as well as the establishment of the clinical prognosis and the initiation of an adequate therapeutic approach. The miniexon PCR-RFLP assay will facilitate such determination and might improve diagnosis and treatment of leishmaniasis.
Publisher
American Society for Microbiology
Reference44 articles.
1. Leishmania and human immunodeficiency virus coinfection: the first 10 years
2. Banuls, A. L., M. Hide, and M. Tibayrenc. 1999. Molecular epidemiology and evolutionary genetics of Leishmania parasites. Int. J. Parasitol.29:1137-1147.
3. Berman, J. D. 1997. Human leishmaniasis: clinical, diagnostic, and chemotherapeutic developments in the last 10 years. Clin. Infect. Dis.24:684-703.
4. Bezold, G., M. Lange, K. Gethoffer, H. Pillekamp, H. Reindl, C. Richter, G. Schonian, L. Weber, and R. U. Peter. 2001. Competitive polymerase chain reaction used to diagnose cutaneous leishmaniasis in German soldiers infected during military exercises in French Guiana. Eur. J. Clin. Microbiol. Infect. Dis.20:421-424.
5. Bhattacharyya, R., K. Das, S. Sen, S. Roy, and H. K. Majumder. 1996. Development of a genus specific primer set for detection of Leishmania parasites by polymerase chain reaction. FEMS Microbiol. Lett.135:195-200.
Cited by
165 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献