An Assessment of Whole Blood and Fractions by Nested PCR as a DNA Source for Diagnosing Canine Ehrlichiosis and Anaplasmosis
Ehrlichiosis and anaplasmosis are tick-borne diseases. Ehrlichia canis and Anaplasma platys infect mainly white cells and platelets, respectively. The main DNA source for PCR is peripheral blood, but the potential of blood cell fractions has not been extensively investigated. This study aims at asse...
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| Language: | English |
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Wiley
2012-01-01
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| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1100/2012/605743 |
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| author | Tereza Emmanuelle de Farias Rotondano Alzira Maria Paiva de Almeida Elane Maria Camboim Lustosa Aline Antas Cordeiro Expedito Kennedy Alves Camboim Sérgio Santos de Azevedo Paulo Paes de Andrade Marcia Almeida de Melo |
| author_facet | Tereza Emmanuelle de Farias Rotondano Alzira Maria Paiva de Almeida Elane Maria Camboim Lustosa Aline Antas Cordeiro Expedito Kennedy Alves Camboim Sérgio Santos de Azevedo Paulo Paes de Andrade Marcia Almeida de Melo |
| author_sort | Tereza Emmanuelle de Farias Rotondano |
| collection | DOAJ |
| description | Ehrlichiosis and anaplasmosis are tick-borne diseases. Ehrlichia canis and Anaplasma platys infect mainly white cells and platelets, respectively. The main DNA source for PCR is peripheral blood, but the potential of blood cell fractions has not been extensively investigated. This study aims at assessment of whole blood (WB) and blood fractions potential in nested PCR (nPCR) to diagnose canine ehrlichiosis and anaplasmosis. The 16S rRNA gene was amplified in 71.4, 17.8, 31.57, and 30% of the WB, granulocyte (G), mononuclear cells (M), and buffy coat (BC) samples. Compared to the WB, the sensitivity of the PCR was 42.86% for the M, and BC fractions, 21.43% for the G, and 33.33% for the blood clot (C). There was fair agreement between the WB and M, BC and C, and slight with the G. Fair agreement occurred between the nPCR and morulae in the blood smear. One animal was coinfected with A. platys and E. canis. This study provided the first evidence of A. platys infection in dogs in Paraíba, Brazil, and demonstrated that WB is a better DNA source than blood fractions to detect Ehrlichia and Anaplasma by nPCR, probably because of the plasma bacterial concentration following host cell lysis. |
| format | Article |
| id | doaj-art-2db209f5c15b4790aec038e746996677 |
| institution | DOAJ |
| issn | 1537-744X |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | The Scientific World Journal |
| spelling | doaj-art-2db209f5c15b4790aec038e7469966772025-08-20T03:20:26ZengWileyThe Scientific World Journal1537-744X2012-01-01201210.1100/2012/605743605743An Assessment of Whole Blood and Fractions by Nested PCR as a DNA Source for Diagnosing Canine Ehrlichiosis and AnaplasmosisTereza Emmanuelle de Farias Rotondano0Alzira Maria Paiva de Almeida1Elane Maria Camboim Lustosa2Aline Antas Cordeiro3Expedito Kennedy Alves Camboim4Sérgio Santos de Azevedo5Paulo Paes de Andrade6Marcia Almeida de Melo7Departamento de Ciências Biológicas, Universidade Federal de Pernambuco, Avenue Professor Moraes Rego, s/n, Cidade Universitária, 50.670-901 Recife, PE, BrazilCentro de Pesquisas Aggeu Magalhães, FIOCRUZ-PE, Avenue Professor Moraes Rego, s/n, Cidade Universitária, 50.670-901 Recife, PE, BrazilUnidade Acadêmica de Medicina Veterinária, Universidade Federal de Campina Grande, 58.700-970 Patos, PB, BrazilUnidade Acadêmica de Medicina Veterinária, Universidade Federal de Campina Grande, 58.700-970 Patos, PB, BrazilUnidade Acadêmica de Medicina Veterinária, Universidade Federal de Campina Grande, 58.700-970 Patos, PB, BrazilUnidade Acadêmica de Medicina Veterinária, Universidade Federal de Campina Grande, 58.700-970 Patos, PB, BrazilDepartamento de Genética, Universidade Federal de Pernambuco, Avenue Professor Moraes Rego, s/n, Cidade Universitária, 50.670-901 Recife, PE, BrazilUnidade Acadêmica de Medicina Veterinária, Universidade Federal de Campina Grande, 58.700-970 Patos, PB, BrazilEhrlichiosis and anaplasmosis are tick-borne diseases. Ehrlichia canis and Anaplasma platys infect mainly white cells and platelets, respectively. The main DNA source for PCR is peripheral blood, but the potential of blood cell fractions has not been extensively investigated. This study aims at assessment of whole blood (WB) and blood fractions potential in nested PCR (nPCR) to diagnose canine ehrlichiosis and anaplasmosis. The 16S rRNA gene was amplified in 71.4, 17.8, 31.57, and 30% of the WB, granulocyte (G), mononuclear cells (M), and buffy coat (BC) samples. Compared to the WB, the sensitivity of the PCR was 42.86% for the M, and BC fractions, 21.43% for the G, and 33.33% for the blood clot (C). There was fair agreement between the WB and M, BC and C, and slight with the G. Fair agreement occurred between the nPCR and morulae in the blood smear. One animal was coinfected with A. platys and E. canis. This study provided the first evidence of A. platys infection in dogs in Paraíba, Brazil, and demonstrated that WB is a better DNA source than blood fractions to detect Ehrlichia and Anaplasma by nPCR, probably because of the plasma bacterial concentration following host cell lysis.http://dx.doi.org/10.1100/2012/605743 |
| spellingShingle | Tereza Emmanuelle de Farias Rotondano Alzira Maria Paiva de Almeida Elane Maria Camboim Lustosa Aline Antas Cordeiro Expedito Kennedy Alves Camboim Sérgio Santos de Azevedo Paulo Paes de Andrade Marcia Almeida de Melo An Assessment of Whole Blood and Fractions by Nested PCR as a DNA Source for Diagnosing Canine Ehrlichiosis and Anaplasmosis The Scientific World Journal |
| title | An Assessment of Whole Blood and Fractions by Nested PCR as a DNA Source for Diagnosing Canine Ehrlichiosis and Anaplasmosis |
| title_full | An Assessment of Whole Blood and Fractions by Nested PCR as a DNA Source for Diagnosing Canine Ehrlichiosis and Anaplasmosis |
| title_fullStr | An Assessment of Whole Blood and Fractions by Nested PCR as a DNA Source for Diagnosing Canine Ehrlichiosis and Anaplasmosis |
| title_full_unstemmed | An Assessment of Whole Blood and Fractions by Nested PCR as a DNA Source for Diagnosing Canine Ehrlichiosis and Anaplasmosis |
| title_short | An Assessment of Whole Blood and Fractions by Nested PCR as a DNA Source for Diagnosing Canine Ehrlichiosis and Anaplasmosis |
| title_sort | assessment of whole blood and fractions by nested pcr as a dna source for diagnosing canine ehrlichiosis and anaplasmosis |
| url | http://dx.doi.org/10.1100/2012/605743 |
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