title: A DNA sequence capture extraction method for detection of Mycobacterium avium subspecies paratuberculosis in feces and tissue samples.
abstract: Culturing of Mycobacterium avium subspecies paratuberculosis (Map) remains difficult and is time consuming. An alternative for the rapid detection of Map in samples is PCR. We have developed a sensitive DNA-extraction method based on sequence capture for the rapid detection of M. avium subspecies paratuberculosis by PCR in fecal and tissue samples. The method detected 10(2)Map/g feces using spiked samples, and reached a diagnostic sensitivity of 33,7% compared to 22% for culture. Analysis of tissue samples gave 65 polymerase chain reaction (PCR)-positive (42.2%) and 49 culture-positive samples (31.8%). Therefore, the detection limit of the DNA-extraction is the same as previously reported for culture, the PCR assay could detect more positive samples than the culture method.
authors.affiliation: Institute of Tropical Medicine, Department of Animal Health, Nationalestraat 155, 2000 Antwerp, Belgium.
authors.forename: Elke
authors.initials: E
authors.lastname: Vansnick
authors.forename: Pim
authors.initials: P
authors.lastname: de Rijk
authors.forename: Francis
authors.initials: F
authors.lastname: Vercammen
authors.forename: Leen
authors.initials: L
authors.lastname: Rigouts
authors.forename: Françoise
authors.initials: F
authors.lastname: Portaels
authors.forename: Dirk
authors.initials: D
authors.lastname: Geysen
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D003670:Deer / Q000382:microbiology; D005243:Feces / Q000382:microbiology*; D007422:Intestines / Q000382:microbiology; D008198:Lymph Nodes / Q000382:microbiology; D016927:Mycobacterium avium subsp. paratuberculosis / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D010283:Paratuberculosis / Q000175:diagnosis* / Q000382:microbiology*; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D004269:DNA, Bacterial