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Identification of Leishmania Species Isolated from Human Cutaneous Leishmaniasis in Mehran, Western Iran Using Nested PCR.

Feiz Haddad MH, Ghasemi E, Maraghi S, Tavala M - Iran J Parasitol (2016 Jan-Mar)

Bottom Line: About 56.3% of patients had a single lesion and a significant correlation was observed between age and number of lesions (P > 0.05).The nested PCR technique was shown to be an effective method with high sensitivity and specificity for identification of human Leishmania parasites.Molecular analysis revealed that parasites isolated from Mehran were identified as L. major and the disease was rural in form.

View Article: PubMed Central - PubMed

Affiliation: Health Research Institute, Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; Dept. of Parasitology, Faculty of Medical Sciences, Ahvaz University of Medical Sciences, Ahvaz, Iran.

ABSTRACT

Background: The incidence of cutaneous leishmaniasis in the city of Mehran has risen sharply in recent years because the city borders Iraq, which has allowed entrance of different Leishmania strains. These strains have different shapes, periods of disease, and healing of lesions. The present study identified and determined cutaneous leishmaniasis species in this region.

Methods: This cross-sectional study was carried out by preparing slides from 92 patients with suspected cutaneous leishmaniasis lesions from Mehran during 2012-2013. Parasite genomic DNA was extracted and CSB2XF and CSB1XR primers were used to amplify the Leishmania minicircle kDNA regions. The parasite species were detected by specific 13Z and LIR primers by applying nested PCR technique.

Results: All banding patterns were diagnosed as L. major parasite by comparison of standard models with amplified fragments 560 bp in length from bands. The patients were 56.5% male and 43.5% female. The most frequently-infected age group was the 21-30 years group at a rate of 27.2%. About 56.3% of patients had a single lesion and a significant correlation was observed between age and number of lesions (P > 0.05).

Conclusion: The nested PCR technique was shown to be an effective method with high sensitivity and specificity for identification of human Leishmania parasites. Molecular analysis revealed that parasites isolated from Mehran were identified as L. major and the disease was rural in form.

No MeSH data available.


Related in: MedlinePlus

Agarose gel electrophoresis of Leishmania isolates. Lane 1, DNA size marker 100 bp; lane 2, negative control; lane3, L. major (positive control 560 bp); lanes 4, 5, 6, 7 and 8, L. major isolates obtained from skin lesions of the patients in Mehran
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Figure 1: Agarose gel electrophoresis of Leishmania isolates. Lane 1, DNA size marker 100 bp; lane 2, negative control; lane3, L. major (positive control 560 bp); lanes 4, 5, 6, 7 and 8, L. major isolates obtained from skin lesions of the patients in Mehran

Mentions: The Leishmania bodies of the 92 isolates were detected by microscopic observation. The samples were all tested by PCR technique. Electrophoresis of the nested PCR products and comparison of the banding patterns with those of the standard samples showed that the bands of all 92 isolates were 560 bp in length, which is standard for L. major (Fig. 1).


Identification of Leishmania Species Isolated from Human Cutaneous Leishmaniasis in Mehran, Western Iran Using Nested PCR.

Feiz Haddad MH, Ghasemi E, Maraghi S, Tavala M - Iran J Parasitol (2016 Jan-Mar)

Agarose gel electrophoresis of Leishmania isolates. Lane 1, DNA size marker 100 bp; lane 2, negative control; lane3, L. major (positive control 560 bp); lanes 4, 5, 6, 7 and 8, L. major isolates obtained from skin lesions of the patients in Mehran
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4835471&req=5

Figure 1: Agarose gel electrophoresis of Leishmania isolates. Lane 1, DNA size marker 100 bp; lane 2, negative control; lane3, L. major (positive control 560 bp); lanes 4, 5, 6, 7 and 8, L. major isolates obtained from skin lesions of the patients in Mehran
Mentions: The Leishmania bodies of the 92 isolates were detected by microscopic observation. The samples were all tested by PCR technique. Electrophoresis of the nested PCR products and comparison of the banding patterns with those of the standard samples showed that the bands of all 92 isolates were 560 bp in length, which is standard for L. major (Fig. 1).

Bottom Line: About 56.3% of patients had a single lesion and a significant correlation was observed between age and number of lesions (P > 0.05).The nested PCR technique was shown to be an effective method with high sensitivity and specificity for identification of human Leishmania parasites.Molecular analysis revealed that parasites isolated from Mehran were identified as L. major and the disease was rural in form.

View Article: PubMed Central - PubMed

Affiliation: Health Research Institute, Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; Dept. of Parasitology, Faculty of Medical Sciences, Ahvaz University of Medical Sciences, Ahvaz, Iran.

ABSTRACT

Background: The incidence of cutaneous leishmaniasis in the city of Mehran has risen sharply in recent years because the city borders Iraq, which has allowed entrance of different Leishmania strains. These strains have different shapes, periods of disease, and healing of lesions. The present study identified and determined cutaneous leishmaniasis species in this region.

Methods: This cross-sectional study was carried out by preparing slides from 92 patients with suspected cutaneous leishmaniasis lesions from Mehran during 2012-2013. Parasite genomic DNA was extracted and CSB2XF and CSB1XR primers were used to amplify the Leishmania minicircle kDNA regions. The parasite species were detected by specific 13Z and LIR primers by applying nested PCR technique.

Results: All banding patterns were diagnosed as L. major parasite by comparison of standard models with amplified fragments 560 bp in length from bands. The patients were 56.5% male and 43.5% female. The most frequently-infected age group was the 21-30 years group at a rate of 27.2%. About 56.3% of patients had a single lesion and a significant correlation was observed between age and number of lesions (P > 0.05).

Conclusion: The nested PCR technique was shown to be an effective method with high sensitivity and specificity for identification of human Leishmania parasites. Molecular analysis revealed that parasites isolated from Mehran were identified as L. major and the disease was rural in form.

No MeSH data available.


Related in: MedlinePlus