DNA amplification assay for rapid detection of bovine tubercle bacilli in semen

被引:6
作者
Ahmed, ASN
Khan, JR
Ganai, NA
机构
[1] CTR DNA Fingerprinting & Diagnost, Hyderabad 500007, Andhra Pradesh, India
[2] Coll Vet Sci, Durg 491001, MP, India
[3] Natl Dairy Res Inst, Karnal 132001, Haryana, India
关键词
cattle-male reproduction; semen characteristics; DNA amplification; Mycobacterium tuberculosis; insertion sequence IS 1081;
D O I
10.1016/S0378-4320(99)00053-6
中图分类号
S8 [畜牧、 动物医学、狩猎、蚕、蜂];
学科分类号
0905 ;
摘要
Tubercle bacilli shed in the semen can be a potential hazard for unlimited number of cows through artificial insemination. We have evaluated the efficacy of a DNA amplification technique by polymerase chain reaction (PCR) for the detection of tubercle bacilli in fresh and frozen semen using spiked samples, The test was based on insertion sequence IS 1081 and could detect as low as 10-100 bacterial cells per mi of spiked semen. The specificity of the test was 100%. The method was applied to semen samples from known and suspected tuberculous bulls. Each of 20 semen samples (fresh and frozen diluted) from one of the three breeding bulls included in the study was found positive while the remaining 40 samples from the other two bulls failed to generate any detectable signal. PCR products were confirmed with Southern blot hybridization to an alpha P-32 labeled-PCR product of the target sequence from the IS 1081 element of the Mycobacterium tuberculosis complex. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:15 / 21
页数:7
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