Comparison of PCR methods for diagnosis of canine visceral leishmaniasis in conjunctival swab samples
Four PCR assays for detection of Leishmania DNA in conjunctival swab samples were compared. All methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or seminested). Two methods (kDNA PCR-hybridization and kDNA snPCR) used primers targeted to the mi...
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| Veröffentlicht in: | Research in veterinary science 2009-10, Vol.87 (2), p.255-257 |
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| description | Four PCR assays for detection of Leishmania DNA in conjunctival swab samples were compared. All methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or seminested). Two methods (kDNA PCR-hybridization and kDNA snPCR) used primers targeted to the minicircles of kinetoplast DNA (kDNA) and the other two methods to the coding (LnPCR) and intergenic noncoding regions (ITS-1 nPCR) of ribosomal rRNA genes. kDNA PCR-hybridization was positive for 22/23 dogs (95.6%) and for 40/46 samples (86.9%), considering the right and the left conjunctivas. kDNA snPCR was positive for 21/23 dogs (91.3%) and for 40/46 samples (86.9%). The ITS-1 nPCR and LnPCR were both able to detect the parasites in 17/23 dogs (73.9%) and 29/46 (63%) and 30/46 (65.2%) samples, respectively. The positivities of the kDNA based methods were significantly higher; however the choice of the best method will depend on the kind of information required with the diagnosis. |
| doi_str_mv | 10.1016/j.rvsc.2009.02.005 |
| format | Article |
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All methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or seminested). Two methods (kDNA PCR-hybridization and kDNA snPCR) used primers targeted to the minicircles of kinetoplast DNA (kDNA) and the other two methods to the coding (LnPCR) and intergenic noncoding regions (ITS-1 nPCR) of ribosomal rRNA genes. kDNA PCR-hybridization was positive for 22/23 dogs (95.6%) and for 40/46 samples (86.9%), considering the right and the left conjunctivas. kDNA snPCR was positive for 21/23 dogs (91.3%) and for 40/46 samples (86.9%). The ITS-1 nPCR and LnPCR were both able to detect the parasites in 17/23 dogs (73.9%) and 29/46 (63%) and 30/46 (65.2%) samples, respectively. The positivities of the kDNA based methods were significantly higher; however the choice of the best method will depend on the kind of information required with the diagnosis.</description><identifier>ISSN: 0034-5288</identifier><identifier>EISSN: 1532-2661</identifier><identifier>DOI: 10.1016/j.rvsc.2009.02.005</identifier><identifier>PMID: 19298988</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>accuracy ; Animals ; Canine visceral leishmaniasis ; conjunctiva ; Conjunctival swab ; Deoxyribonucleic acid ; Diagnosis ; diagnostic techniques ; disease detection ; disease diagnosis ; DNA ; DNA Primers ; DNA, Protozoan - genetics ; DNA, Protozoan - isolation & purification ; dog diseases ; Dog Diseases - diagnosis ; Dog Diseases - parasitology ; Dogs ; Gene Amplification ; Genes, Protozoan - genetics ; Leishmania ; Leishmania - genetics ; Leishmaniasis, Visceral - diagnosis ; Leishmaniasis, Visceral - genetics ; Leishmaniasis, Visceral - veterinary ; Methods ; microbial genetics ; Microbiology ; molecular sequence data ; Nucleic Acid Hybridization ; nucleotide sequences ; Parasites ; Parasitic diseases ; pathogen identification ; polymerase chain reaction ; Polymerase chain reaction (PCR) ; Polymerase Chain Reaction - methods ; RNA, Protozoan - genetics ; RNA, Protozoan - isolation & purification ; RNA, Ribosomal - genetics ; RNA, Ribosomal - isolation & purification ; Veterinary medicine ; visceral leishmaniasis</subject><ispartof>Research in veterinary science, 2009-10, Vol.87 (2), p.255-257</ispartof><rights>2009 Elsevier Ltd</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c437t-a07fd4d2c76ad4a86d601c749e5600f576c1a6868d015a21f74cfc09d3253d413</citedby><cites>FETCH-LOGICAL-c437t-a07fd4d2c76ad4a86d601c749e5600f576c1a6868d015a21f74cfc09d3253d413</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.rvsc.2009.02.005$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19298988$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pilatti, Marcia Maria</creatorcontrib><creatorcontrib>Ferreira, Sidney de Almeida</creatorcontrib><creatorcontrib>Melo, Maria Norma de</creatorcontrib><creatorcontrib>Andrade, Antero Silva Ribeiro de</creatorcontrib><title>Comparison of PCR methods for diagnosis of canine visceral leishmaniasis in conjunctival swab samples</title><title>Research in veterinary science</title><addtitle>Res Vet Sci</addtitle><description>Four PCR assays for detection of Leishmania DNA in conjunctival swab samples were compared. All methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or seminested). Two methods (kDNA PCR-hybridization and kDNA snPCR) used primers targeted to the minicircles of kinetoplast DNA (kDNA) and the other two methods to the coding (LnPCR) and intergenic noncoding regions (ITS-1 nPCR) of ribosomal rRNA genes. kDNA PCR-hybridization was positive for 22/23 dogs (95.6%) and for 40/46 samples (86.9%), considering the right and the left conjunctivas. kDNA snPCR was positive for 21/23 dogs (91.3%) and for 40/46 samples (86.9%). The ITS-1 nPCR and LnPCR were both able to detect the parasites in 17/23 dogs (73.9%) and 29/46 (63%) and 30/46 (65.2%) samples, respectively. The positivities of the kDNA based methods were significantly higher; however the choice of the best method will depend on the kind of information required with the diagnosis.</description><subject>accuracy</subject><subject>Animals</subject><subject>Canine visceral leishmaniasis</subject><subject>conjunctiva</subject><subject>Conjunctival swab</subject><subject>Deoxyribonucleic acid</subject><subject>Diagnosis</subject><subject>diagnostic techniques</subject><subject>disease detection</subject><subject>disease diagnosis</subject><subject>DNA</subject><subject>DNA Primers</subject><subject>DNA, Protozoan - genetics</subject><subject>DNA, Protozoan - isolation & purification</subject><subject>dog diseases</subject><subject>Dog Diseases - diagnosis</subject><subject>Dog Diseases - parasitology</subject><subject>Dogs</subject><subject>Gene Amplification</subject><subject>Genes, Protozoan - genetics</subject><subject>Leishmania</subject><subject>Leishmania - genetics</subject><subject>Leishmaniasis, Visceral - diagnosis</subject><subject>Leishmaniasis, Visceral - genetics</subject><subject>Leishmaniasis, Visceral - veterinary</subject><subject>Methods</subject><subject>microbial genetics</subject><subject>Microbiology</subject><subject>molecular sequence data</subject><subject>Nucleic Acid Hybridization</subject><subject>nucleotide sequences</subject><subject>Parasites</subject><subject>Parasitic diseases</subject><subject>pathogen identification</subject><subject>polymerase chain reaction</subject><subject>Polymerase chain reaction (PCR)</subject><subject>Polymerase Chain Reaction - methods</subject><subject>RNA, Protozoan - genetics</subject><subject>RNA, Protozoan - isolation & purification</subject><subject>RNA, Ribosomal - genetics</subject><subject>RNA, Ribosomal - isolation & purification</subject><subject>Veterinary medicine</subject><subject>visceral leishmaniasis</subject><issn>0034-5288</issn><issn>1532-2661</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkV2L1TAQhoMo7nH1D3ihBcG71knSpC14sxz8ggVF3euQzcduSpscM-0R_70p54DghV4NzDzzMslDyHMKDQUq34xNPqJpGMDQAGsAxAOyo4KzmklJH5IdAG9rwfr-gjxBHAGgpbR7TC7owIZ-6Psdcfs0H3QOmGKVfPVl_7Wa3XKfLFY-5coGfRcTBtyGRscQXXUMaFzWUzW5gPdzaeoNCLEyKY5rNEs4lin-1LcV6vkwOXxKHnk9oXt2rpfk5v277_uP9fXnD5_2V9e1aXm31Bo6b1vLTCe1bXUvrQRqunZwQgJ40UlDtexlb4EKzajvWuMNDJYzwW1L-SV5fco95PRjdbioeTt2mnR0aUUlOwGMD_8H2YYNdCjgq7_AMa05lkcoClxAW0RscexEmZwQs_PqkMOs868Cqc2VGtXmSm2uFDBVXJWlF-fo9XZ29s_KWU4BXp4Ar5PSd0WSuvnGgPISKJlkG_H2RLjyqcfgskITXDTOhuzMomwK_7rgNzksrkQ</recordid><startdate>20091001</startdate><enddate>20091001</enddate><creator>Pilatti, Marcia Maria</creator><creator>Ferreira, Sidney de Almeida</creator><creator>Melo, Maria Norma de</creator><creator>Andrade, Antero Silva Ribeiro de</creator><general>Elsevier Ltd</general><general>Elsevier Limited</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QG</scope><scope>7QP</scope><scope>7QR</scope><scope>7T5</scope><scope>7T7</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U7</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>K9.</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20091001</creationdate><title>Comparison of PCR methods for diagnosis of canine visceral leishmaniasis in conjunctival swab samples</title><author>Pilatti, Marcia Maria ; Ferreira, Sidney de Almeida ; Melo, Maria Norma de ; Andrade, Antero Silva Ribeiro de</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c437t-a07fd4d2c76ad4a86d601c749e5600f576c1a6868d015a21f74cfc09d3253d413</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>accuracy</topic><topic>Animals</topic><topic>Canine visceral leishmaniasis</topic><topic>conjunctiva</topic><topic>Conjunctival swab</topic><topic>Deoxyribonucleic acid</topic><topic>Diagnosis</topic><topic>diagnostic techniques</topic><topic>disease detection</topic><topic>disease diagnosis</topic><topic>DNA</topic><topic>DNA Primers</topic><topic>DNA, Protozoan - genetics</topic><topic>DNA, Protozoan - isolation & purification</topic><topic>dog diseases</topic><topic>Dog Diseases - diagnosis</topic><topic>Dog Diseases - parasitology</topic><topic>Dogs</topic><topic>Gene Amplification</topic><topic>Genes, Protozoan - genetics</topic><topic>Leishmania</topic><topic>Leishmania - genetics</topic><topic>Leishmaniasis, Visceral - diagnosis</topic><topic>Leishmaniasis, Visceral - genetics</topic><topic>Leishmaniasis, Visceral - veterinary</topic><topic>Methods</topic><topic>microbial genetics</topic><topic>Microbiology</topic><topic>molecular sequence data</topic><topic>Nucleic Acid Hybridization</topic><topic>nucleotide sequences</topic><topic>Parasites</topic><topic>Parasitic diseases</topic><topic>pathogen identification</topic><topic>polymerase chain reaction</topic><topic>Polymerase chain reaction (PCR)</topic><topic>Polymerase Chain Reaction - methods</topic><topic>RNA, Protozoan - genetics</topic><topic>RNA, Protozoan - isolation & purification</topic><topic>RNA, Ribosomal - genetics</topic><topic>RNA, Ribosomal - isolation & purification</topic><topic>Veterinary medicine</topic><topic>visceral leishmaniasis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pilatti, Marcia Maria</creatorcontrib><creatorcontrib>Ferreira, Sidney de Almeida</creatorcontrib><creatorcontrib>Melo, Maria Norma de</creatorcontrib><creatorcontrib>Andrade, Antero Silva Ribeiro de</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Immunology Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Research in veterinary science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pilatti, Marcia Maria</au><au>Ferreira, Sidney de Almeida</au><au>Melo, Maria Norma de</au><au>Andrade, Antero Silva Ribeiro de</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparison of PCR methods for diagnosis of canine visceral leishmaniasis in conjunctival swab samples</atitle><jtitle>Research in veterinary science</jtitle><addtitle>Res Vet Sci</addtitle><date>2009-10-01</date><risdate>2009</risdate><volume>87</volume><issue>2</issue><spage>255</spage><epage>257</epage><pages>255-257</pages><issn>0034-5288</issn><eissn>1532-2661</eissn><abstract>Four PCR assays for detection of Leishmania DNA in conjunctival swab samples were compared. All methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or seminested). Two methods (kDNA PCR-hybridization and kDNA snPCR) used primers targeted to the minicircles of kinetoplast DNA (kDNA) and the other two methods to the coding (LnPCR) and intergenic noncoding regions (ITS-1 nPCR) of ribosomal rRNA genes. kDNA PCR-hybridization was positive for 22/23 dogs (95.6%) and for 40/46 samples (86.9%), considering the right and the left conjunctivas. kDNA snPCR was positive for 21/23 dogs (91.3%) and for 40/46 samples (86.9%). The ITS-1 nPCR and LnPCR were both able to detect the parasites in 17/23 dogs (73.9%) and 29/46 (63%) and 30/46 (65.2%) samples, respectively. The positivities of the kDNA based methods were significantly higher; however the choice of the best method will depend on the kind of information required with the diagnosis.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>19298988</pmid><doi>10.1016/j.rvsc.2009.02.005</doi><tpages>3</tpages></addata></record> |
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| subjects | accuracy Animals Canine visceral leishmaniasis conjunctiva Conjunctival swab Deoxyribonucleic acid Diagnosis diagnostic techniques disease detection disease diagnosis DNA DNA Primers DNA, Protozoan - genetics DNA, Protozoan - isolation & purification dog diseases Dog Diseases - diagnosis Dog Diseases - parasitology Dogs Gene Amplification Genes, Protozoan - genetics Leishmania Leishmania - genetics Leishmaniasis, Visceral - diagnosis Leishmaniasis, Visceral - genetics Leishmaniasis, Visceral - veterinary Methods microbial genetics Microbiology molecular sequence data Nucleic Acid Hybridization nucleotide sequences Parasites Parasitic diseases pathogen identification polymerase chain reaction Polymerase chain reaction (PCR) Polymerase Chain Reaction - methods RNA, Protozoan - genetics RNA, Protozoan - isolation & purification RNA, Ribosomal - genetics RNA, Ribosomal - isolation & purification Veterinary medicine visceral leishmaniasis |
| title | Comparison of PCR methods for diagnosis of canine visceral leishmaniasis in conjunctival swab samples |
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