Hamdy Mahmoud E R, Amin A S
Department of Agricultural Development, Animal Health Affairs, Doha, State of Qatar, Egypt.
Vet J. 2002 May;163(3):299-305. doi: 10.1053/tvjl.2001.0681.
One hundred and three milk samples were collected from 52 cows, 21 ewes, 18 goats and 12 camels. The animals tested positive to at least one of the following: (1) standard tube agglutination test (SAT); (2) Rose Bengal plate test (RBPT); (3) milk ring test (MRT). All milk samples were examined by culture and single-step polymerase chain reaction (PCR) techniques for detection of Brucella species. The PCR assay amplified Brucella-DNA from 29 bovine milk samples, 10 from sheep, 13 from goats and one from a camel. The direct culture method detected Brucella organisms from 24 samples of cows' milk, 12 from sheep, 10 from goats and failed to detect any Brucella organisms from camels' milk. PCR detected up to 100 colony forming units (CFU) of B. abortus per millilitre of milk in 100% of diluted milk samples, and 1000 CFU of B. melitensis from 70% of milk samples. Although the overall sensitivity of the PCR was higher than the culture method, it should be possible to increase the sensitivity to detect lower numbers of Brucella organisms in field samples. The speed and sensitivity of the PCR assay suggest that this technique could be useful for detection of Brucella organisms in bovine milk, as well as in sheep, goat, and camels milk.
从52头奶牛、21只母羊、18只山羊和12头骆驼身上采集了103份牛奶样本。这些动物对以下至少一项检测呈阳性：(1) 标准试管凝集试验（SAT）；(2) 玫瑰红平板试验（RBPT）；(3) 牛奶环状试验（MRT）。所有牛奶样本均通过培养和单步聚合酶链反应（PCR）技术检测布鲁氏菌属菌种。PCR检测从29份牛奶样本中扩增出布鲁氏菌DNA，其中10份来自绵羊，13份来自山羊，1份来自骆驼。直接培养法从24份牛奶样本中检测到布鲁氏菌，12份来自绵羊，10份来自山羊，而从骆驼奶中未检测到任何布鲁氏菌。PCR在100%的稀释牛奶样本中检测到每毫升牛奶中含有高达100个流产布鲁氏菌菌落形成单位（CFU），在70% 的牛奶样本中检测到每毫升含有1000个马尔他布鲁氏菌CFU。尽管PCR的总体灵敏度高于培养法，但应该有可能提高灵敏度以检测野外样本中数量更少 的布鲁氏菌。PCR检测的速度和灵敏度表明，该技术可用于检测牛奶中的布鲁氏菌，以及绵羊奶、山羊奶和骆驼奶中的布鲁氏菌。
