Identification of Leishmania Species Using PCR Assay on Giemsa-Stained Slides Prepared From Cutaneous Leishmaniasis Patients

Background Leishmaniasis is a group of diseases that are created by intracellular parasites of Leishmania. Cutaneous leishmaniasis is considered as one of the health problems in some provinces of Iran. Methods In this study, a total of 178 Giemsa-stained slides from confirmed cases of cutaneous leishmaniasis were examined. The slides were prepared from the patients with cutaneous leishmaniasis that referred to health centers and infected during the epidemic of cutaneous leishmaniasis in Poldokhtar city, Lorestan Province, Iran in 2006.Genomic DNA from each slide was extracted. After DNA extraction, ITS-PCR was used. Results Out of 178 slides, 129 (72.47%) samples had a band in the range of 485 bp and 49 (27.53%) samples 626 bp that matched L. tropica and L. major standard samples, respectively. Conclusion This study showed that Leishmania DNA could be efficiently extracted and amplified even from old Giemsa-stained microscopic slides that were stored more than 6 yr. In this study was shown that both L. tropica and L. major species exist in Lorestan Province.

[1]  A. Khamesipour,et al.  First molecular identification of Leishmania species in a new endemic area of cutaneous leishmaniasis in Lorestan, Iran. , 2013, Asian Pacific journal of tropical medicine.

[2]  I. Sharifi,et al.  Spatial Distribution and Molecular Identification of Leishmania Species from Endemic Foci of South-Eastern Iran , 2012, Iranian journal of parasitology.

[3]  J. Saki,et al.  Diagnosis and identification of Leishmania spp. from Giemsa-stained slides, by real-time PCR and melting curve analysis in south-west of Iran , 2011, Annals of tropical medicine and parasitology.

[4]  İ. Koltaş,et al.  Identification of Causative Species in Cutaneous Leishmaniasis Patients Using PCR-RFLP , 2011 .

[5]  Oshaghi Mohammad Ali,et al.  Phlebotomus papatasi and Meriones libycus as the vector and reservoir host of cutaneous leishmaniasis in Qomrood District, Qom Province, central Iran. , 2011, Asian Pacific journal of tropical medicine.

[6]  H. Nahrevanian,et al.  An overview of a diagnostic and epidemiologic reappraisal of cutaneous leishmaniasis in Iran. , 2011, The Brazilian journal of infectious diseases : an official publication of the Brazilian Society of Infectious Diseases.

[7]  E. Nasiri,et al.  EPIDEMIOLOGICAL SURVEY OF CUTANEOUS LEISHMANIASIS IN LORESTAN PROVINCE (IRAN) AND INTRODUCTION OF DISEASE TRANSMISSION IN NEW LOCAL AREAS , 2011 .

[8]  S. Spalding,et al.  Molecular techniques for the study and diagnosis of parasite infection , 2011 .

[9]  H. Al-Mekhlafi,et al.  Molecular Characterization of Leishmania Species Isolated from Cutaneous Leishmaniasis in Yemen , 2010, PloS one.

[10]  Suljey Cochero,et al.  Molecular identification of the parasites causing cutaneous leishmaniasis on the Caribbean coast of Colombia , 2010, Parasitology Research.

[11]  J. Jafari,et al.  Some Epidemiological Aspects of Cutaneous Leishmaniasis in Northern Khorasan Province, Iran , 2009, Iranian journal of arthropod-borne diseases.

[12]  M. Mahmoudi Rad,et al.  Identification of leishmania species isolated from human cutaneous leishmaniasis using PCR method. , 2009, Journal of research in health sciences.

[13]  M. Baghaei,et al.  A new focus of zoonotic cutaneous leishmaniasis in Shiraz, Iran. , 2009, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[14]  J. Sadraei,et al.  Determination of Leishmania species causing cutaneous leishmaniasis in Mashhad by PCR-RFLP method , 2009 .

[15]  Theresa L. Smith,et al.  Chapter 5 – Other Infectious Diseases Related to Travel , 2009 .

[16]  M. Mohebali,et al.  Molecular Identification of Leishmania Species Isolated from Human Cutaneous Leishmaniasis by RAPD-PCR , 2009 .

[17]  B. Kazemi,et al.  Evaluation of PCR assay in diagnosis and identification of cutaneous leishmaniasis: a comparison with the parasitological methods , 2008, Parasitology Research.

[18]  M. Mohebali,et al.  Diagnosis and characterization of Leishmania species in Giemsa-stained slides by PCR-RFLP. , 2008 .

[19]  P. Karanis,et al.  Evaluation of PCR-RFLP (based on ITS-1 and HaeIII) for the detection of Leishmania species, using Greek canine isolates and Jordanian clinical material , 2007, Annals of tropical medicine and parasitology.

[20]  C. Pirmez,et al.  Sensitivity and specificity of polymerase chain reaction in Giemsa-stained slides for diagnosis of visceral leishmaniasis in children. , 2007, Memorias do Instituto Oswaldo Cruz.

[21]  M. F. Araújo,et al.  PCR identification of Leishmania in diagnosis and control of canine Leishmaniasis. , 2007, Veterinary parasitology.

[22]  S. Maraghi,et al.  Identification of Cutaneous Leishmaniasis Agents by Nested Po- lymerase Chain Reaction (Nested-PCR) in Shush City, Khuzestan Province, Iran , 2007 .

[23]  F. Prugnolle,et al.  Leishmania and the leishmaniases: a parasite genetic update and advances in taxonomy, epidemiology and pathogenicity in humans. , 2007, Advances in parasitology.

[24]  A. Volpini,et al.  Leishmania identification by PCR of Giemsa-stained lesion imprint slides stored for up to 36 years. , 2006, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[25]  G. Schoenian,et al.  Clinical diagnosis of cutaneous leishmaniasis: a comparison study between standardized graded direct microscopy and ITS1-PCR of Giemsa-stained smears. , 2006, Acta tropica.

[26]  H. Vatandoost,et al.  Meriones libycus is the main reservoir of zoonotic cutaneous leishmaniasis in south Islamic Republic of Iran. , 2006, Eastern Mediterranean health journal = La revue de sante de la Mediterranee orientale = al-Majallah al-sihhiyah li-sharq al-mutawassit.

[27]  Sarman Singh New developments in diagnosis of leishmaniasis. , 2006, The Indian journal of medical research.

[28]  C. Jaffe,et al.  PCR diagnosis and characterization of Leishmania in local and imported clinical samples. , 2003, Diagnostic microbiology and infectious disease.

[29]  C. Jaffe,et al.  Identification and Differentiation of Leishmania Species in Clinical Samples by PCR Amplification of the Miniexon Sequence and Subsequent Restriction Fragment Length Polymorphism Analysis , 2003, Journal of Clinical Microbiology.

[30]  H. Noyes,et al.  DNA extraction and amplification of Leishmania from archived, Giemsa-stained slides, for the diagnosis of cutaneous leishmaniasis by PCR , 2002, Annals of tropical medicine and parasitology.

[31]  C J Marinkelle,et al.  The control of leishmaniases. , 1980, Bulletin of the World Health Organization.