A DNA sequence capture extraction method for detection of Mycobacterium avium subspecies paratuberculosis in feces and tissue samples.
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.
Vansnick
Elke
E
Institute of Tropical Medicine, Department of Animal Health, Nationalestraat 155, 2000 Antwerp, Belgium.
de Rijk
Pim
P
Vercammen
Francis
F
Rigouts
Leen
L
Portaels
Françoise
F
Geysen
Dirk
D
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*
D004269:DNA, Bacterial