Comparison of different tissue sampling for PCR-based diagnosis and follow-up of canine visceral leishmaniosis.
In this study, different types of tissue sampling for PCR-based diagnosis and follow-up of canine visceral leishmaniosis were compared. Skin, whole blood and lymph node samples were collected from 95 naturally infected dogs living in South Italy, where the disease is endemic. Twenty-nine of these 95 dogs, treated with meglumine administered concurrently with allopurinol for 30 days, and then with allopurinol alone, were monitored during a period of 2 years. The DNA extracted from the clinical specimens was amplified by PCR using as target DNA a 116-bp fragment in the constant region of the kinetoplast DNA minicircle. PCR analysis was more sensitive than indirect immunofluorescence antibody test in detecting Leishmania infection in symptomatic dogs: 99% of lymph node samples resulted positive, whereas 94% of blood samples and 95% of skin samples gave a positive result. PCR analysis of samples from dogs followed up 2 years showed that: (1) all subjects resulted positive in at least one of the three types of samples; (2) all time the dogs had a relapse, PCR resulted positive in all three types of samples; (3) when dogs were apparently healthy, PCR analysis was positive on skin and lymph node samples, but not always on blood samples. Since lymph node sampling is invasive and sometimes difficult in healthy asymptomatic dogs, our results suggest that, independently from the presence or not of cutaneous lesions, skin biopsy represents a good substratum for PCR-based diagnosis and follow-up of canine visceral leishmaniosis.
Manna
Laura
L
Dipartimento di Scienze Cliniche Veterinarie, Università di Napoli Federico II, Via F. Delpino 1, 80137 Naples, Italy.
Vitale
Fabrizio
F
Reale
Stefano
S
Caracappa
Santo
S
Pavone
Luigi Michele
LM
Morte
Rossella Della
RD
Cringoli
Giuseppe
G
Staiano
Norma
N
Gravino
Angelo Elio
AE
D000493:Allopurinol / Q000627:therapeutic use; D000818:Animals; D000981:Antiprotozoal Agents / Q000627:therapeutic use; D001706:Biopsy / Q000662:veterinary; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004283:Dog Diseases / Q000175:diagnosis / Q000188:drug therapy / Q000469:parasitology*; D004285:Dogs; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D018314:Leishmania infantum / Q000235:genetics / Q000302:isolation & purification*; D007898:Leishmaniasis, Visceral / Q000175:diagnosis / Q000188:drug therapy / Q000469:parasitology / Q000662:veterinary*; D008198:Lymph Nodes / Q000469:parasitology; D008297:Male; D008536:Meglumine / Q000627:therapeutic use; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D012876:Skin Diseases, Parasitic / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*
D000981:Antiprotozoal Agents; D004272:DNA, Mitochondrial; D016054:DNA, Protozoan; D000493:Allopurinol; D008536:Meglumine