Detection of Mycobacterium avium subsp. paratuberculosis in bovine fecal samples: comparison of three polymerase chain reaction-based diagnostic tests with a conventional culture method.
Three commercially available assays, designed to specifically detect the presence of Mycobacterium avium subsp. paratuberculosis (MAP) in fecal samples by IS900-PCR, were compared with a conventional culture method. Fecal samples from 100 dairy cows were tested. Fifty-four (67.5%) of 80 culture-positive samples were positive for an assay that detects MAP DNA by dot spot hybridization of polymerase chain reaction products (kit A), 48 (60%) were positive by an assay using ethidium bromide staining for agar gel visualization of amplification products (kit B), and 49 (61.3%) were positive by an assay in which amplified products are detected by a colorimetric detection system (kit C). Relative sensitivity of all tests increased in proportion to the presence of MAP in fecal samples. Specificity was 100% based on results from 20 culture-negative samples from an MAP-free herd.
Taddei
Simone
S
Sezione di Malattie Infettive degli Animali, Dipartimento di Salute Animale, Università di Parma, 43100 Parma, Italy.
Robbi
Chiara
C
Cesena
Camilla
C
Rossi
Ilenia
I
Schiano
Emiliana
E
Arrigoni
Norma
N
Vicenzoni
Gaddo
G
Cavirani
Sandro
S
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000382:microbiology*; D003124:Colorimetry / Q000662:veterinary; D004269:DNA, Bacterial / Q000032:analysis; D005243:Feces / Q000382:microbiology*; D016927:Mycobacterium avium subsp. paratuberculosis / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D010283:Paratuberculosis / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*
D004269:DNA, Bacterial