Cutaneous Leishmaniasis in El-Madinna Manowra region, Saudi Arabia in 2012 (original) (raw)
Related papers
Annals of Tropical Medicine & Parasitology, 2011
In Iran, Leishmania major or L. tropica cause almost all of the human cutaneous leishmaniasis (CL). Unfortunately, the detection methods frequently used for CL (the microscopical examination of direct smears or the culture of biopsies) are not very sensitive and the Leishmania species causing each case of CL in Iran is usually only tentatively identified from extrinsic factors, such as the case's clinical manifestations and region of residence. Recently, however, a nested PCR that targets the parasites' kinetoplast DNA has been used in the city of Ahvaz (the capital of the province of Khouzestan, in southwestern Iran) to confirm the microscopical diagnosis of CL and to identify the causative parasites, to species level. Smears from the lesions on 100 suspected cases of CL were fixed, stained with Wright's eosin-methylene blue, and checked for amastigotes under a light microscope. Scrapings from the same smears were then tested for leishmanial DNA, using a nested PCR that allows the DNA from L. tropica to be identified and distinguished from that of L. major. The 100 smears investigated were all found amastigote-positive by microscopy and PCR-positive for either L. major DNA (97 smears) or L. tropica DNA (three smears). The predominant species causing CL in Ahvaz is therefore L. major.
Identification of the causative agent of cutaneous leishmaniasis in Chichaoua province, Morocco
Parasite, 2012
Cutaneous leishmaniasis (CL) in Morocco is caused by three species, Leishmania major, L. tropica and L. infantum. CL has been known in Chichaoua province since 2000. Using DNA extracted from microscopic slides and parasite cultures, collected in the years 2006 and 2009, we identified for the first time L. tropica as the causative agent of CL in this region. Species identification was achieved by performing the ITS1-PCR-RFLP approach. By using this method it was possible to identify parasites in Giemsa stained slides containing less than five parasites per oil-immersion field even they were conserved for up to four months. KEY WORDS: leishmaniasis, Leishmania tropica, ITS1-PCR, Morocco. Résumé : IDENTIFICATION DE L'AGENT CAUSAL DE LA LEISHMANIOSE CUTANÉE DANS LA PROVINCE DE CHICHAOUA, AU MAROC La leishmaniose cutanée (LC) dans la province de Chichaoua est connue depuis l'an 2000. Cette étude montre pour la première fois l'espèce parasitaire circulante dans cette province. L'identité du parasite a été déterminée sur des lames colorées et des cultures de parasites en utilisant l'ITS1-PCR, suivie de la digestion par l'enzyme HaeIII. Le résultat a montré que Leishmania tropica est l'agent responsable de la LC. Par ailleurs, une corrélation entre le nombre de parasites par lame et la positivité du test moléculaire a été réalisée et a montré que l'ITS1-PCR permet d'identifier des parasites dans les lames colorées au Giemsa contenant moins de cinq parasites par champs et même si elles étaient conservées jusqu'à quatre mois.
Identification of Species Causing Cutaneous Leishmaniasis by PCR in Chahbahar, Iran
Medical Laboratory Journal, 2016
Background and Objective: Chabahar is in Southern Iran located near the Iran-Pakistan border. Since leishmaniasis is an emerging disease in this region, this study aimed to diagnose the disease and identify different species of Leishmania parasite in the patients referred to the central laboratory. Methods: This descriptive cross-sectional study was conducted in 2011-2012 on patients referred to the central laboratory in the city of Chabahar. The sampling of lesions, slide preparation, culture and PCR specific for kinetoplast DNA (kDNA), extracted from the media and slides, were performed. The data collected by a questionnaire were analyzed by the SPSS software. Results: The resulted bands from the 48 tested cutaneous leishmaniasis isolates were compared with the standard strains of Leishmania tropica, L. infantum and L. major. All 48 investigated bands were in the 620bp region, which is related to L. major. Conclusion: Since PCR has high sensitivity and specificity, it is recommended to use kDNA (present in a unique organelle called kinetoplast) for the routine diagnosis and treatment of the disease.
International Journal of Infectious Diseases, 2013
Background: Leishmaniasis is a parasitic disease affecting a large number of people worldwide. In this study we carried out the molecular characterization of cutaneous leishmaniasis (CL) in Al-Madinah Al-Munawarah Province, Saudi Arabia, confirming Leishmania major and Leishmania tropica as the prevalent species using molecular techniques. Methods: One hundred and five patients with suspected CL were identified from four different localities in Al-Madinah Al-Munawarah Province and Al-Miqat Hospital, Al-Madinah, Saudi Arabia. Thirty-four of the 105 patients were selected at random for molecular investigation. Results: Characterization of CL species by internal transcribed spacer 1 (ITS1) PCR-restriction fragment length polymorphism (RFLP) and kinetoplast DNA (kDNA) PCR established L. major and L. tropica as the causative organisms. kDNA PCR had a sensitivity of 90.7%, whereas ITS1 PCR had a sensitivity of 70.1%, thus facilitating the diagnosis and species identification. Parasite culture alone detected 39.2% and smear alone 55.3% of the positive samples. With the exception of kDNA PCR, all other assays were 100% specific. Conclusions: This study provides the first findings for the comprehensive molecular characterization of CL in Saudi Arabia.
Background: The diagnosis of cutaneous leishmaniasis (CL) might be difficult, in particular in endemic areas where different species of Leishmania can cause lesions of very similar appearance and where other skin diseases with similar clinical symptoms occur. Even today, the parasitological diagnosis of CL remains the gold standard and it is based on the direct identification of amastigotes in microscopy smears and/or culture of promastigotes from infected tissues. Although these techniques are highly specific, they are not sensitive enough. The objective of this study is to contribute to improving the diagnosis of CL and the identification of Leishmania species in Morocco by comparing three PCR-based assays applied directly on dermal samples. Methods: A total of 58 patients presenting with cutaneous lesions suggestive of CL were sampled for parasitological diagnosis by direct examination (DE), culture in NNN medium, two kinetoplast DNA (kDNA) PCRs (Lmj4/Uni21 and 13A/13B primers) and one rRNA gene internal transcribed spacer 1 (ITS1) PCR (LITSR/L5.8S primers). The techniques were statistically analyzed and compared.
Jundishapur Journal of Microbiology, 2013
Background: Leishmaniasis is a common parasitic disease world wide. Leishmania tropica and L. major are two common cause of cutaneous leishmaniasis in Iran. The aim of this study was determination of the cause of cutaneous leishmaniasis in Shush city, Khouzestan Province, Southwest Iran Methods: One hundred samples were collected from patients at the age of 1-80 year with documented cutaneous leishmaniasis referred to the health centre and a private medical diagnostic laboratory at Shush City. DNA was extracted from slid samples by phenol-chloroform-Isoemil alcohol method, and subjected to Nested-PCR as template. k DNA of the parasites were amplified by CSB1XR and CSB2XF in the first round of PCR and 13Z and Li R primers for the second round. After PCR, electrophoresis of products was performed and 750bp band from L. tropica and 560bp band from L. major were detected. Results: A total of 100 cases comprising 47 females and 53 males were studied. The highest infected age group was under 10 years with a rate of 42% and the lowest rate was 4% at the age group of above 40 years. The results of PCR electrophoresis indicated that 90(90%) cases were L. major and 10 (10%) L. tropica. The predominant species in this area was L. major. Conclusion: It is concluded that Nested PCR is a reliable test for diagnosis and identification of Leishmania species and can apply in epidemiological investigations.
Usefulness of PCR in the diagnosis of cutaneous leishmaniasis in Tunisia
Transactions of the Royal Society of Tropical Medicine and Hygiene, 2005
We assessed the efficiency of a PCR method in establishing the diagnosis of cutaneous leishmaniasis (CL) in Tunisian patients. Four hundred and thirty specimens collected passively from patients with cutaneous ulcers suggestive of leishmaniasis attending health centres for diagnosis were included in the study. Dermal scrapings were analysed both by parasitological (examination of Giemsa-stained smears and in vitro cultivation) methods and by a genus-specific PCR detecting a fragment of the 18S rRNA gene. Microscopy revealed amastigotes in 245 samples (57.0%) and in vitro cultivation gave positive results in 88 cases (20.5%), whereas PCR detected Leishmania in 301 samples (70%). The sensitivities inferred from our results were 99.3%, 80.8% and 29% for PCR, microscopic examination and in vitro cultivation, respectively. The different forms of CL in this country are caused by three species of Leishmania and are treated with the same protocol. Of 303 welldocumented cases in our study, 99% were probably caused by Leishmania major and 1% by Leishmania infantum. The lack of species-specific diagnosis is not known to affect treatment or prognosis in Tunisia. These data support the incorporation of PCR into diagnostic strategies for CL, particularly in Tunisia.
Parasitology Research, 2008
The aims of this study are to identify Leishmania species and compare and validate internal transcribed spacers (ITS) polymerase chain reaction (PCR) assay against parasitological methods for diagnosing cutaneous leishmaniasis (CL). We used the ITS-PCR, parasite culture, and microscopic evaluation of stained smears on 155 specimens from suspected cases of (CL) patients who referred to Mashhad Health Centers (northeast Iran). The PCR indicated the sensitivity (98.8%), correctly diagnosing 86 of the 87 confirmed positive specimens. Microscopy and parasite culture alone showed 79.3% sensitivity (69/87 positive) and 86.2% sensitivity (75/87 positive), respectively, while microscopy and culture in combination improved sensitivity totally to 100% (87/87). The results also revealed that Leishmania tropica species is dominant (96.5%) in the studied regions. This study suggests that both the parasitological techniques reliably were used for the diagnosis of CL, and the ITS-PCR assay without using RFLP analysis is useful for identifying Leishmania species in the area.
Molecular identification of Leishmania species causing cutaneous leishmaniasis in Mashhad, Iran
2010
Background: Cutaneous Leishmaniasis (CL) is a parasitic skin disease. Diagnosis primarily is based on clinical signs and microscopic observation of parasite on direct stained smears or tissue sections. Sensitivity of direct smear is not as high as molecular methods. The aim of this study was to identify and characterize Leishmania species among the negative direct smears obtained from skin ulcers suspected to CL by PCR method. Methods: Among 81 patients with suspicious skin lesions to CL referred to the Parasitology lab, negative Giemsa stained smears were collected. DNA extraction performed by scraping stained smears, then PCR was performed. Results: Among the DNA extracted from smears, L. tropica was isolated from 9 (11.1%) of the smears and L.major was not isolated from any samples. Conclusion: Direct microscopy on stained smears for diagnosis of leishmaniasis is not enough accurate. PCR is recommended for clinically suspected lesions with negative result of direct smear.