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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Bibliographic Details
Published in:Research in veterinary science 2009-10, Vol.87 (2), p.255-257
Main Authors: Pilatti, Marcia Maria, Ferreira, Sidney de Almeida, Melo, Maria Norma de, Andrade, Antero Silva Ribeiro de
Format: Article
Language:English
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
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Summary: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.
ISSN:0034-5288
1532-2661
DOI:10.1016/j.rvsc.2009.02.005