Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis
Objective: The objective of this study was to analyze different primers that are commonly used in epidemiological studies for the detection of Leishmania DNA by PCR, and to compare them to the conventional direct parasite search for American cutaneous leishmaniasis (ACL) diagnosis. Material and meth...
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| Format: | Article |
| Language: | English |
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Elsevier
2011-05-01
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| Series: | Brazilian Journal of Infectious Diseases |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1413867011701763 |
| _version_ | 1818309878283763712 |
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| author | Diego Molina de Oliveira, M.D. Maria Valdrinez Campana Lonardoni, M.D.Ph.D. Ueslei Teodoro Thais Gomes Verzignassi Silveira |
| author_facet | Diego Molina de Oliveira, M.D. Maria Valdrinez Campana Lonardoni, M.D.Ph.D. Ueslei Teodoro Thais Gomes Verzignassi Silveira |
| author_sort | Diego Molina de Oliveira, M.D. |
| collection | DOAJ |
| description | Objective: The objective of this study was to analyze different primers that are commonly used in epidemiological studies for the detection of Leishmania DNA by PCR, and to compare them to the conventional direct parasite search for American cutaneous leishmaniasis (ACL) diagnosis. Material and methods: Five pairs of primers, four of them derived from Leishmania kDNA sequences (MP3H-MP1L; B1-B2; LBF1-LBR1; 13A-13B), and one derived from the SL RNA (mini-exon) gene repeat (LU5A-LB3C), reported previously, were used Results and discussion: The MP3H-MP1L primers were the best at amplifying the DNA, detecting 2 fg of Leishmania spp. DNA. The 13A-13B primers presented the worst performance, detecting 512 x 103 fg of DNA. Conclusion: The wide variation in the analytical sensitivity of the primers used in the PCR, and the significant differences from the conventional method of ACL diagnosis found in this study, emphasize the importance of standardizing the PCR technique, analyzing sensitivity, and selecting suitable oligonucleotide primers. Keywords: leishmaniasis, polymerase chain reaction, Leishmania, DNA primers |
| first_indexed | 2024-12-13T07:37:10Z |
| format | Article |
| id | doaj.art-48190f2252df4223a1425b6f2c27801d |
| institution | Directory Open Access Journal |
| issn | 1413-8670 |
| language | English |
| last_indexed | 2024-12-13T07:37:10Z |
| publishDate | 2011-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Brazilian Journal of Infectious Diseases |
| spelling | doaj.art-48190f2252df4223a1425b6f2c27801d2022-12-21T23:55:03ZengElsevierBrazilian Journal of Infectious Diseases1413-86702011-05-01153204210Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasisDiego Molina de Oliveira, M.D.0Maria Valdrinez Campana Lonardoni, M.D.Ph.D.1Ueslei Teodoro2Thais Gomes Verzignassi Silveira3Postgraduate Student, Universidade Estadual de Maringá, PR, BrazilDepartament of Clinical Analysis, Universidade Estadual de Maringá, PR, BrazilDepartament of Clinical Analysis, Universidade Estadual de Maringá, PR, BrazilDepartament of Clinical Analysis, Universidade Estadual de Maringá, PR, Brazil; Departamento de Análises Clínicas e Biomedicina, Universidade Estadual de Maringá, Av. Colombo 5790, Maringá, Paraná, Brazil 87020-900.Objective: The objective of this study was to analyze different primers that are commonly used in epidemiological studies for the detection of Leishmania DNA by PCR, and to compare them to the conventional direct parasite search for American cutaneous leishmaniasis (ACL) diagnosis. Material and methods: Five pairs of primers, four of them derived from Leishmania kDNA sequences (MP3H-MP1L; B1-B2; LBF1-LBR1; 13A-13B), and one derived from the SL RNA (mini-exon) gene repeat (LU5A-LB3C), reported previously, were used Results and discussion: The MP3H-MP1L primers were the best at amplifying the DNA, detecting 2 fg of Leishmania spp. DNA. The 13A-13B primers presented the worst performance, detecting 512 x 103 fg of DNA. Conclusion: The wide variation in the analytical sensitivity of the primers used in the PCR, and the significant differences from the conventional method of ACL diagnosis found in this study, emphasize the importance of standardizing the PCR technique, analyzing sensitivity, and selecting suitable oligonucleotide primers. Keywords: leishmaniasis, polymerase chain reaction, Leishmania, DNA primershttp://www.sciencedirect.com/science/article/pii/S1413867011701763 |
| spellingShingle | Diego Molina de Oliveira, M.D. Maria Valdrinez Campana Lonardoni, M.D.Ph.D. Ueslei Teodoro Thais Gomes Verzignassi Silveira Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis Brazilian Journal of Infectious Diseases |
| title | Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis |
| title_full | Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis |
| title_fullStr | Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis |
| title_full_unstemmed | Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis |
| title_short | Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis |
| title_sort | comparison of different primes for pcr based diagnosis of cutaneous leishmaniasis |
| url | http://www.sciencedirect.com/science/article/pii/S1413867011701763 |
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