Quantification of Mycobacterium avium subsp paratuberculosis (MAP) survival in monocyte-derived macrophages

被引：6

作者：

Mitchell, Rebecca M. ^{[1
]}

Gollnick, Nicole S. ^{[2
]}

Sreevatsan, Srinand ^{[3
]}

Russell, David G. ^{[4
]}

Schukken, Ynte H. ^{[1
]}

机构：

[1] Cornell Univ, Coll Vet Med, Dept Populat Med & Diagnost Sci, Ithaca, NY 14853 USA

[2] Univ Munich, Clin Ruminants, D-85764 Oberschleissheim, Germany

[3] Univ Minnesota, Coll Vet Med, Dept Vet Populat Med, St Paul, MN 55108 USA

[4] Cornell Univ, Coll Vet Med, Dept Microbiol & Immunol, Ithaca, NY 14853 USA

来源：

VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY | 2011年 / 139卷 / 01期

关键词：

Mycobacterium avium subsp; paratuberculosis; PBMC; Quantitative PCR; Fluorescence; DIVERSITY; CATTLE;

D O I：

10.1016/j.vetimm.2010.08.003

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

Real-time PCR assays were developed to quantitate Mycobacterium avium subsp. paratuberculosis (MAP) in bovine monocyte-derived macrophages. We measured the absolute number of both host cells and bacteria in in vitro challenge assays. Results obtained from real-time quantitative PCR (qPCR) DNA copy counts were compared to visual quantitation of fluorescent-stained MAP in macrophages. Conclusions from our original visual analysis were supported by the second (qPCR) methodology; however, the qPCR assay proved to be more consistent between samples and was easier to perform. There was a strain-to-strain difference in growth curves between fluorescent quantitation (FQ) and qPCR that we believe to be a consequence of bacterial growth characteristics in FQ. In summary, real-time PCR assays provided a more accurate and precise method for evaluating intracellular growth dynamics when comparing strains of MAP. (C) 2010 Elsevier B.V. All rights reserved.

引用

页码：73 / 78

页数：6

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