Comparison of Three PCR-based Methods for Simplicity and Cost Effectiveness Identification of Cutaneous Leishmaniasis Due to Leishmania tropica
Background: To compare three molecular methods, PCR-RFLP for internal transcribed spacer, PCR sequencing and high resolution melting analysis shown reliable sensitivity and specificity for detecting Leishmania tropica as a model for cutaneous leishmaniasis (CL) as the perspective overview for scient...
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2017-06-01
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| Series: | Iranian Journal of Parasitology |
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| Online Access: | https://ijpa.tums.ac.ir/index.php/ijpa/article/view/1628 |
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| author | Mohammad Ali MOHAMMADI Mehdi BAMOROVAT Majid FASIHI HARANDI Tayyebeh KARIMI Iraj SHARIFI Mohammad Reza AFLATOONIAN |
| author_facet | Mohammad Ali MOHAMMADI Mehdi BAMOROVAT Majid FASIHI HARANDI Tayyebeh KARIMI Iraj SHARIFI Mohammad Reza AFLATOONIAN |
| author_sort | Mohammad Ali MOHAMMADI |
| collection | DOAJ |
| description | Background: To compare three molecular methods, PCR-RFLP for internal transcribed spacer, PCR sequencing and high resolution melting analysis shown reliable sensitivity and specificity for detecting Leishmania tropica as a model for cutaneous leishmaniasis (CL) as the perspective overview for scientific and economic approaches.
Methods: This study was carried out between 2015 and 2016 in Leishmaniasis Research Center in Kerman University of Medical Sciences, Kerman, Iran. The positives smears (n=50) were obtained from patients referred from the health clinics in a major anthroponotic CL (ACL) focus, southeastern Iran. Only smear preparations with the same grade were selected according to the method described by the WHO for future PCR assays.
Results: All three molecular methods had capability to identify positive samples at species level with the same specificity and sensitivity. However, these techniques were different in simplicity, consuming time, and cost effectiveness. Although additional enzymatic process in PCR-RFLP provided good resolution to find Leishmania species but this would cause time and cost increases.
Conclusion: HRM (high resolution melting) is a relatively new technique that allows direct characterization of PCR amplicons in a closed system with more simplicity, cost effectiveness and time-consuming compared with other PCR-based assays for epidemiological or clinical identification purposes. |
| first_indexed | 2024-04-11T14:30:57Z |
| format | Article |
| id | doaj.art-78d0c12b1b23403da98ce523e3538de6 |
| institution | Directory Open Access Journal |
| issn | 1735-7020 2008-238X |
| language | English |
| last_indexed | 2024-04-11T14:30:57Z |
| publishDate | 2017-06-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Parasitology |
| spelling | doaj.art-78d0c12b1b23403da98ce523e3538de62022-12-22T04:18:38ZengTehran University of Medical SciencesIranian Journal of Parasitology1735-70202008-238X2017-06-01122656Comparison of Three PCR-based Methods for Simplicity and Cost Effectiveness Identification of Cutaneous Leishmaniasis Due to Leishmania tropicaMohammad Ali MOHAMMADI0Mehdi BAMOROVAT1Majid FASIHI HARANDI2Tayyebeh KARIMI3Iraj SHARIFI4Mohammad Reza AFLATOONIAN5Research Center for Hydatid Disease in Iran, Kerman University of Medical Sciences, Kerman, IranLeishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, IranLeishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, IranDept. of Medical Parasitology, School of Medicine, Kerman University of Medical Sciences, Kerman, IranLeishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, IranResearch Center for Tropical and Infectious Diseases, Kerman University of Medical Sciences, Kerman, IranBackground: To compare three molecular methods, PCR-RFLP for internal transcribed spacer, PCR sequencing and high resolution melting analysis shown reliable sensitivity and specificity for detecting Leishmania tropica as a model for cutaneous leishmaniasis (CL) as the perspective overview for scientific and economic approaches. Methods: This study was carried out between 2015 and 2016 in Leishmaniasis Research Center in Kerman University of Medical Sciences, Kerman, Iran. The positives smears (n=50) were obtained from patients referred from the health clinics in a major anthroponotic CL (ACL) focus, southeastern Iran. Only smear preparations with the same grade were selected according to the method described by the WHO for future PCR assays. Results: All three molecular methods had capability to identify positive samples at species level with the same specificity and sensitivity. However, these techniques were different in simplicity, consuming time, and cost effectiveness. Although additional enzymatic process in PCR-RFLP provided good resolution to find Leishmania species but this would cause time and cost increases. Conclusion: HRM (high resolution melting) is a relatively new technique that allows direct characterization of PCR amplicons in a closed system with more simplicity, cost effectiveness and time-consuming compared with other PCR-based assays for epidemiological or clinical identification purposes.https://ijpa.tums.ac.ir/index.php/ijpa/article/view/1628Molecular identificationCutaneous leishmania-sisPCRIran |
| spellingShingle | Mohammad Ali MOHAMMADI Mehdi BAMOROVAT Majid FASIHI HARANDI Tayyebeh KARIMI Iraj SHARIFI Mohammad Reza AFLATOONIAN Comparison of Three PCR-based Methods for Simplicity and Cost Effectiveness Identification of Cutaneous Leishmaniasis Due to Leishmania tropica Iranian Journal of Parasitology Molecular identification Cutaneous leishmania-sis PCR Iran |
| title | Comparison of Three PCR-based Methods for Simplicity and Cost Effectiveness Identification of Cutaneous Leishmaniasis Due to Leishmania tropica |
| title_full | Comparison of Three PCR-based Methods for Simplicity and Cost Effectiveness Identification of Cutaneous Leishmaniasis Due to Leishmania tropica |
| title_fullStr | Comparison of Three PCR-based Methods for Simplicity and Cost Effectiveness Identification of Cutaneous Leishmaniasis Due to Leishmania tropica |
| title_full_unstemmed | Comparison of Three PCR-based Methods for Simplicity and Cost Effectiveness Identification of Cutaneous Leishmaniasis Due to Leishmania tropica |
| title_short | Comparison of Three PCR-based Methods for Simplicity and Cost Effectiveness Identification of Cutaneous Leishmaniasis Due to Leishmania tropica |
| title_sort | comparison of three pcr based methods for simplicity and cost effectiveness identification of cutaneous leishmaniasis due to leishmania tropica |
| topic | Molecular identification Cutaneous leishmania-sis PCR Iran |
| url | https://ijpa.tums.ac.ir/index.php/ijpa/article/view/1628 |
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