Diagnosis of Cutaneous Leishmaniasis and Species Discrimination of Parasites by PCR and Hybridization
The aim of this study was to assess the efficacy of PCR methodology in establishing the diagnosis of cutaneous leishmaniasis in patients from areas of endemicity in Venezuela. Biopsies from 233 patients with cutaneous ulcers suggestive of leishmaniasis were analyzed by PCR, employing oligonucleotide...
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1994
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| Online Access: | http://hdl.handle.net/10872/12694 |
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| _version_ | 1870621935928344576 |
|---|---|
| bitstream.checksum.fl_str_mv | 9874ac9db136cab8fd5fa73f2b572d42 7c020282d630887ba819753d2e4a4c58 |
| bitstream.checksumAlgorithm.fl_str_mv | MD5 MD5 |
| bitstream.url.fl_str_mv | https://saber.ucv.ve/jspui/bitstream/10872/12694/1/Rodriguez%20y%20Takiff.pdf https://saber.ucv.ve/jspui/bitstream/10872/12694/2/license.txt |
| dc.contributor.author.none.fl_str_mv | Rodríguez, Noris Guzman, Bernardo Rodas, A. Takiff, Howard R.Bloom, Barry Convit, Jacinto |
| dc.creator.fl_str_mv | Rodríguez, Noris Guzman, Bernardo Rodas, A. Takiff, Howard R.Bloom, Barry Convit, Jacinto |
| dc.date.accessioned.none.fl_str_mv | 2015-11-09T20:16:13Z |
| dc.date.available.none.fl_str_mv | 2015-11-09T20:16:13Z |
| dc.date.issued.none.fl_str_mv | 1994-09 |
| dc.identifier.issn.none.fl_str_mv | 2246–2252. |
| dc.identifier.uri.none.fl_str_mv | http://hdl.handle.net/10872/12694 |
| dc.language.iso.es_VE.fl_str_mv | en |
| dc.publisher.es_VE.fl_str_mv | JOURNAL OF CLINICAL MICROBIOLOGY |
| dc.relation.ispartofseries.none.fl_str_mv | Vol.32;9 |
| dc.subject.es_VE.fl_str_mv | Diagnosis Hybridization Leishmaniasis DISCRIMINATION |
| dc.title.es_VE.fl_str_mv | Diagnosis of Cutaneous Leishmaniasis and Species Discrimination of Parasites by PCR and Hybridization |
| dc.type.es_VE.fl_str_mv | Article |
| description | The aim of this study was to assess the efficacy of PCR methodology in establishing the diagnosis of cutaneous leishmaniasis in patients from areas of endemicity in Venezuela. Biopsies from 233 patients with cutaneous ulcers suggestive of leishmaniasis were analyzed by PCR, employing oligonucleotides directed against conserved regions of kinetoplast DNA (kDNA), and the PCR products were then hybridized to nonradioactively labeled, species-specific, cloned kDNA fragments. The ability of PCR to detect Leishmania cells was compared with those of the conventional methodologies: skin testing with killed promastigotes (Montenegro test), examination of Giemsa-stained biopsy smears, and in vitro culture of biopsy tissue. The PCR-hybridization technique detected the presence of Leishmania cells in 98% of patients clinically diagnosed as having leishmaniasis and also positive by the Montenegro skin test. In comparison, leishmania positivity was found in only 42% of cultures and 64% of biopsy smears. By hybridizing the PCR product to new kDNA probes specific for either Leishmania mexicana or Leishmania braziliensis, we found that both species are major causes of cutaneous leishmaniasis in Venezuela, and the species identification was confirmed by restriction enzyme analysis of kDNA from biopsy cultures. This work demonstrates that PCR coupled with hybridization is useful not only for the diagnosis of cutaneous leishmaniasis but also for the taxonomic discrimination essential for both epidemiology and therapy. This technique can be used to diagnose leishmaniasis in a country in which the disease is endemic and can perhaps be adapted for use in a rural clinic. |
| id | SABERUCV_50fd00aef200c2cd2e907ee403848606 |
| identifier_str_mv | 2246–2252. |
| language_invalid_str_mv | en |
| network_acronym_str | SABERUCV |
| network_name_str | Repositorio Institucional de la Universidad Central de Venezuela |
| oai_identifier_str | oai:saber.ucv.ve:10872/12694 |
| publishDate | 1994 |
| publishDateSort | 1994 |
| repository.mail.fl_str_mv | sfwrussians@gmail.com |
| repository.name.fl_str_mv | Saber UCV |
| repository_id_str | |
| spelling | Rodríguez, NorisGuzman, BernardoRodas, A.Takiff, HowardR.Bloom, BarryConvit, Jacinto2015-11-09T20:16:13Z2015-11-09T20:16:13Z1994-092246–2252.http://hdl.handle.net/10872/12694The aim of this study was to assess the efficacy of PCR methodology in establishing the diagnosis of cutaneous leishmaniasis in patients from areas of endemicity in Venezuela. Biopsies from 233 patients with cutaneous ulcers suggestive of leishmaniasis were analyzed by PCR, employing oligonucleotides directed against conserved regions of kinetoplast DNA (kDNA), and the PCR products were then hybridized to nonradioactively labeled, species-specific, cloned kDNA fragments. The ability of PCR to detect Leishmania cells was compared with those of the conventional methodologies: skin testing with killed promastigotes (Montenegro test), examination of Giemsa-stained biopsy smears, and in vitro culture of biopsy tissue. The PCR-hybridization technique detected the presence of Leishmania cells in 98% of patients clinically diagnosed as having leishmaniasis and also positive by the Montenegro skin test. In comparison, leishmania positivity was found in only 42% of cultures and 64% of biopsy smears. By hybridizing the PCR product to new kDNA probes specific for either Leishmania mexicana or Leishmania braziliensis, we found that both species are major causes of cutaneous leishmaniasis in Venezuela, and the species identification was confirmed by restriction enzyme analysis of kDNA from biopsy cultures. This work demonstrates that PCR coupled with hybridization is useful not only for the diagnosis of cutaneous leishmaniasis but also for the taxonomic discrimination essential for both epidemiology and therapy. This technique can be used to diagnose leishmaniasis in a country in which the disease is endemic and can perhaps be adapted for use in a rural clinic.enJOURNAL OF CLINICAL MICROBIOLOGYVol.32;9DiagnosisHybridizationLeishmaniasisDISCRIMINATIONDiagnosis of Cutaneous Leishmaniasis and Species Discrimination of Parasites by PCR and HybridizationArticleORIGINALRodriguez y Takiff.pdfRodriguez y Takiff.pdfapplication/pdf1872229https://saber.ucv.ve/jspui/bitstream/10872/12694/1/Rodriguez%20y%20Takiff.pdf9874ac9db136cab8fd5fa73f2b572d42MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81509https://saber.ucv.ve/jspui/bitstream/10872/12694/2/license.txt7c020282d630887ba819753d2e4a4c58MD5210872/12694oai:saber.ucv.ve:10872/126942015-11-09 20:16:13.94Saber UCVsfwrussians@gmail.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 |
| spellingShingle | Diagnosis of Cutaneous Leishmaniasis and Species Discrimination of Parasites by PCR and Hybridization Diagnosis Hybridization Leishmaniasis DISCRIMINATION |
| title | Diagnosis of Cutaneous Leishmaniasis and Species Discrimination of Parasites by PCR and Hybridization |
| title_full | Diagnosis of Cutaneous Leishmaniasis and Species Discrimination of Parasites by PCR and Hybridization |
| title_fullStr | Diagnosis of Cutaneous Leishmaniasis and Species Discrimination of Parasites by PCR and Hybridization |
| title_full_unstemmed | Diagnosis of Cutaneous Leishmaniasis and Species Discrimination of Parasites by PCR and Hybridization |
| title_short | Diagnosis of Cutaneous Leishmaniasis and Species Discrimination of Parasites by PCR and Hybridization |
| title_sort | Diagnosis of Cutaneous Leishmaniasis and Species Discrimination of Parasites by PCR and Hybridization |
| topic | Diagnosis Hybridization Leishmaniasis DISCRIMINATION |
| url | http://hdl.handle.net/10872/12694 |