Veterinary Laboratories Agency (VLA) - Weybridge, New Haw, Addlestone, Surrey.
Vet Rec. 2010 Dec 18;167(25):965-7. doi: 10.1136/vr.c4040.
A real-time PCR was developed to detect Coxiella burnetii (the cause of Q fever) in ruminant placentas and aborted fetuses. Primer and probe sets previously developed for human tissue studies were used to target the insertion sequence IS1111 gene for C burnetii. The assay was highly sensitive, with a limit of detection of 10 copies of template, theoretically equating to a single bacterium, and did not cross-react with a panel of other bacteria. To determine sensitivity on field samples submitted for the diagnosis of abortion, results using the IS1111 PCR assay were compared with a com1 PCR assay. When applied to ruminant abortion material, including placental cotyledons and fetal samples, the IS1111 and com1 assays yielded positive results in 23 (25 per cent) of 93 and 19 (20 per cent) of 93 samples, respectively. One infected goat herd was monitored for 31 months: 57 (92 per cent) of 62 placental cotyledon samples from aborting and non-aborting goats, and 10 (30 per cent) of 33 fetal samples were positive by the IS1111 PCR assay.
开发了一种实时 PCR 来检测牛布鲁氏菌(引起 Q 热)在反刍动物胎盘中和流产胎儿中的存在。用于人类组织研究的引物和探针集被用于针对插入序列 IS1111 基因检测 C 燃烧菌。该检测方法非常灵敏,检测限为模板的 10 个拷贝,理论上相当于一个单一的细菌,并且不会与其他细菌的面板发生交叉反应。为了确定提交用于诊断流产的现场样本的灵敏度,使用 IS1111 PCR 检测的结果与 com1 PCR 检测进行了比较。当应用于包括胎盘胎叶和胎儿样本的反刍动物流产材料时,IS1111 和 com1 检测分别在 93 个样本中的 23 个(25%)和 93 个样本中的 19 个(20%)呈阳性。对一个受感染的山羊群进行了 31 个月的监测:在流产和非流产山羊的 62 个胎盘胎叶样本中,有 57 个(92%)和 33 个胎儿样本中的 10 个(30%)通过 IS1111 PCR 检测呈阳性。