Genomic DNA extraction and amplification of Leishmania donovani using polymerase chain reaction (PCR) from archived, Giemsa- stained slides
<p>In Sri Lanka, diagnosis of cutaneous leishmaniasis<strong> (</strong>CL) is usually based on clinical features and direct microscopy examinations. Polymerase chain reaction (PCR) for <em>Leishmania</em> is usually performed on tissue samples. In this study, we extrac...
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| Format: | Article |
| Language: | English |
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Sri Lankan Society for Microbiology
2021-04-01
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| Series: | Sri Lankan Journal of Infectious Diseases |
| Subjects: | |
| Online Access: | https://sljid.sljol.info/articles/8323 |
| _version_ | 1797985083416641536 |
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| author | A. Amarasinghe D. Iddawela |
| author_facet | A. Amarasinghe D. Iddawela |
| author_sort | A. Amarasinghe |
| collection | DOAJ |
| description | <p>In Sri Lanka, diagnosis of cutaneous leishmaniasis<strong> (</strong>CL) is usually based on clinical features and direct microscopy examinations. Polymerase chain reaction (PCR) for <em>Leishmania</em> is usually performed on tissue samples. In this study, we extracted DNA from archived Giemsa-stained slides which were prepared from cutaneous lesions. A total number of 85 Giemsa-stained slides fixed between 2008-2017 were selected. All the slides were examined using light microscopy and the number of amastigotes in positive smears was recorded. A nested PCR was carried out to amplify the 385 bp fragment of <em>L. donovani </em>kinetoplast mini-circle sequence. All 40 positive slides had only 1-2 amastigotes per slide. Of these, only 20% were PCR positive. Of the 45 negative slides, only one gave positive PCR result. Further studies are required to confirm the efficacy of PCR on Giemsa-stained smears in our setting.</p> |
| first_indexed | 2024-04-11T07:13:02Z |
| format | Article |
| id | doaj.art-26c43a5a6a3440dbb64b40cfd0d8dc52 |
| institution | Directory Open Access Journal |
| issn | 2012-8169 2448-9654 |
| language | English |
| last_indexed | 2024-04-11T07:13:02Z |
| publishDate | 2021-04-01 |
| publisher | Sri Lankan Society for Microbiology |
| record_format | Article |
| series | Sri Lankan Journal of Infectious Diseases |
| spelling | doaj.art-26c43a5a6a3440dbb64b40cfd0d8dc522022-12-22T04:38:06ZengSri Lankan Society for MicrobiologySri Lankan Journal of Infectious Diseases2012-81692448-96542021-04-01111232610.4038/sljid.v11i1.83235855Genomic DNA extraction and amplification of Leishmania donovani using polymerase chain reaction (PCR) from archived, Giemsa- stained slidesA. Amarasinghe0D. Iddawela1University of Peradeniya, PeradeniyaUniversity of Peradeniya, Peradeniya<p>In Sri Lanka, diagnosis of cutaneous leishmaniasis<strong> (</strong>CL) is usually based on clinical features and direct microscopy examinations. Polymerase chain reaction (PCR) for <em>Leishmania</em> is usually performed on tissue samples. In this study, we extracted DNA from archived Giemsa-stained slides which were prepared from cutaneous lesions. A total number of 85 Giemsa-stained slides fixed between 2008-2017 were selected. All the slides were examined using light microscopy and the number of amastigotes in positive smears was recorded. A nested PCR was carried out to amplify the 385 bp fragment of <em>L. donovani </em>kinetoplast mini-circle sequence. All 40 positive slides had only 1-2 amastigotes per slide. Of these, only 20% were PCR positive. Of the 45 negative slides, only one gave positive PCR result. Further studies are required to confirm the efficacy of PCR on Giemsa-stained smears in our setting.</p>https://sljid.sljol.info/articles/8323cutaneous leishmaniasis, giemsa-stained smears, leishmania donovani, sri lanka |
| spellingShingle | A. Amarasinghe D. Iddawela Genomic DNA extraction and amplification of Leishmania donovani using polymerase chain reaction (PCR) from archived, Giemsa- stained slides Sri Lankan Journal of Infectious Diseases cutaneous leishmaniasis, giemsa-stained smears, leishmania donovani, sri lanka |
| title | Genomic DNA extraction and amplification of Leishmania donovani using polymerase chain reaction (PCR) from archived, Giemsa- stained slides |
| title_full | Genomic DNA extraction and amplification of Leishmania donovani using polymerase chain reaction (PCR) from archived, Giemsa- stained slides |
| title_fullStr | Genomic DNA extraction and amplification of Leishmania donovani using polymerase chain reaction (PCR) from archived, Giemsa- stained slides |
| title_full_unstemmed | Genomic DNA extraction and amplification of Leishmania donovani using polymerase chain reaction (PCR) from archived, Giemsa- stained slides |
| title_short | Genomic DNA extraction and amplification of Leishmania donovani using polymerase chain reaction (PCR) from archived, Giemsa- stained slides |
| title_sort | genomic dna extraction and amplification of leishmania donovani using polymerase chain reaction pcr from archived giemsa stained slides |
| topic | cutaneous leishmaniasis, giemsa-stained smears, leishmania donovani, sri lanka |
| url | https://sljid.sljol.info/articles/8323 |
| work_keys_str_mv | AT aamarasinghe genomicdnaextractionandamplificationofleishmaniadonovaniusingpolymerasechainreactionpcrfromarchivedgiemsastainedslides AT diddawela genomicdnaextractionandamplificationofleishmaniadonovaniusingpolymerasechainreactionpcrfromarchivedgiemsastainedslides |