The detection of Coxiella burnetii from ovine genital swabs, milk and fecal samples by the use of a single touchdown polymerase chain reaction
The detection of Coxiella burnetii from ovine genital swabs, milk and fecal samples by the use of a single touchdown polymerase chain reaction
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DOI：
10.1016/s0378-1135(99)00178-9
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发表时间：
2000-03-15
影响因子：
3.3
通讯作者：
Rodolakis, A
中科院分区：
文献类型：
--
作者：
Berri, M;Laroucau, K;Rodolakis, A
以伯氏柯克斯体重复转座子样区为靶点的聚合酶链式反应(PCR)直接检测绵羊生殖器拭子、乳汁和粪便标本中的伯氏柯克斯体。采用离心、QIAMP组织试剂盒DNA纯化和反式聚合酶链式反应相结合的方法，进一步提高了检测母羊乳样中柯克斯体的效率，并在1毫升乳汁中检出1个伯氏柯克斯体细胞。此外，还介绍了一种有效、简便、快速的方法，只需将DNA模板稀释到1：100，即可从粪便标本中去除PCR抑制物质，从而实现了每毫克样品中一种微生物的检测。从血清阳性的母羊身上获得的结果证明，在自然感染的动物的乳汁和粪便样本中也可以检测到伯氏梭菌。(C)2000 Elsevier Science B.V.保留所有权利。
The polymerase chain reaction (PCR), targeting the repetitive transposon-like region of Coxiella burnetii (Trans-PCR), was evaluated for its ability to detect directly C. burnetii in genital swabs, milk and fecal specimens of ewes. By using a combination of centrifugation step, DNA purification using Qiamp Tissue kit followed by Trans-PCR assay, the efficiency for detection of coxiella in ewes milk samples was further improved and one C. burnetii-cell could be detected in I mi of milk. In addition, an effective, simple and rapid method to remove PCR-inhibitory substances from fecal specimens by simply diluting the DNA template to 1:100 is described, which made the detection of one microorganism per mg of sample then possible. The results obtained from seropositive ewes proved that C. burnetii could also be detected in milk and fecal samples of naturally infected animals. (C) 2000 Elsevier Science B.V. All rights reserved.