Detection of canine distemper virus in dogs by real-time RT-PCR.
Canine distemper virus is the etiological agent of a severe disease in dogs and many other carnivores. Clinical diagnosis of canine distemper is difficult due to the broad spectrum of signs that may be confounded with other respiratory and enteric diseases of dogs. Accordingly, a laboratory confirmation is required for suspected cases. In this study a real-time RT-PCR assay was developed for detection and quantitation of canine distemper virus. The assay exhibited high specificity as all the negative controls (no-template-controls and samples from healthy sero-negative dogs) and other canine pathogens were not misdetected. Up to 1 x 10(2) copies of RNA were detected by the TaqMan assay, thus revealing a high sensitivity. Quantitative TaqMan was validated on clinical samples, including various tissues and organs collected from dogs naturally infected by canine distemper virus. Urines, tonsil, conjunctival swabs and whole blood were found to contain high virus loads and therefore proved to be suitable targets for detection of canine distemper virus RNA.
Elia
Gabriella
G
Department of Animal Health and Well-being, Faculty of Veterinary Medicine of Bari, S.p. per Casamassima km 3, 70010 Valenzano, Bari, Italy. g.elia@veterinaria.uniba.it
Decaro
Nicola
N
Martella
Vito
V
Cirone
Francesco
F
Lucente
Maria Stella
MS
Lorusso
Eleonora
E
Di Trani
Livia
L
Buonavoglia
Canio
C
D000825:Animal Structures / Q000821:virology; D000818:Animals; D001826:Body Fluids / Q000821:virology; D004216:Distemper / Q000175:diagnosis* / Q000821:virology; D004217:Distemper Virus, Canine / Q000235:genetics / Q000302:isolation & purification*; D004285:Dogs; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity