Detection of Mycobacterium avium subspecies paratuberculosis in Swiss dairy cattle by real-time PCR and culture:: a comparison of the two assays

被引:41
作者
Bögli-Stuber, K
Kohler, C
Seitert, G
Glanemann, B
Antognoli, MC
Salman, MD
Wittenbrink, MM
Wittwer, M
Wassenaar, T
Jemmi, T
Bissig-Choisat, B
机构
[1] Swiss Fed Vet Off, Labs, CH-3003 Bern, Switzerland
[2] Univ Zurich, Inst Vet Bacteriol, Zurich, Switzerland
[3] Colorado State Univ, Anim Populat Hlth Inst, Coll Vet Med & Biomed Sci, Ft Collins, CO 80523 USA
[4] Mol Microbiol & Genom Consultants, Zotzenheim, Germany
关键词
culture; DNA extraction; faeces; IS900; real-time PCR;
D O I
10.1111/j.1365-2672.2005.02645.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Aims: To compare the two different diagnostic assays for the detection of Mycobacterium avium ssp. paratuberculosis, the aetiological agent of paratuberculosis. Methods and Results: Faecal samples were derived from 310 cows, representing 13 commercial dairy herds in various locations in Switzerland with expected increased risk because of a past history of disease. Detection assays for M. avium ssp. paratuberculosis were culture (gold standard) and a newly designed real-time PCR. Real-time PCR identified 31 of 310 animals as positive within this risk population whereas culture identified 20 positive animals. The specificity of real-time PCR was confirmed by DNA sequencing of the PCR product. Depending on the test used, the paratuberculosis prevalence in our tested risk population ranged from 6.5 to 10%. Conclusions: Real-time PCR and culture data were in good agreement, and real-time PCR generates data in a short time in contrast to culture. Significance and Impact of the Study: We consider real-time PCR as a suitable alternative method to culture for the detection of M. avium ssp. paratuberculosis in a national surveillance programme.
引用
收藏
页码:587 / 597
页数:11
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