Comparison of polymerase chain reaction assays with bacteriologic culture, immunofluorescence, and nucleic acid hybridization for detection of Leptospira borgpetersenii serovar hardjo in urine of cattle.
Comparison of polymerase chain reaction assays with bacteriologic culture, immunofluorescence, and nucleic acid hybridization for detection of Leptospira borgpetersenii serovar hardjo in urine of cattle.
复制标题
聚合酶链反应检测与细菌学培养、免疫荧光和核酸杂交检测牛尿液中哈德霍钩端螺旋体的比较。
DOI：
10.2460/ajvr.2000.61.316
复制
发表时间：
2000
影响因子：
1
通讯作者：
Carole A. Bolin
中科院分区：
文献类型：
--
作者：
Jaap A. Wagenaar;R. Zuerner;David P. Alt;Carole A. Bolin
目的
比较不同PCR方法检测牛尿中伯氏钩端螺旋体（Leptospira borgpetersenii serovar hardjo）的敏感性和特异性，并将最佳PCR方法的检测结果与免疫荧光法、核酸杂交法和细菌培养法的检测结果进行比较。
动物
六头小母牛。
程序
通过连续3天结膜滴注10（6）个钩端螺旋体，使小母牛暴露于哈德乔型哈德乔牛血清型。在感染前和感染后收集尿液样品。比较了5种PCR检测方法的灵敏度和特异性，以确定用于牛尿液样品的最佳检测方法。然后将优化的PCR检测方法与细菌学培养、核酸杂交和免疫荧光的结果进行比较。
结果
选择特异性（100%）和灵敏度（91%）最佳组合的PCR检测与其他诊断试验进行比较。核酸杂交的敏感性为55%，而细菌培养和免疫荧光的敏感性为89%至93%。
结论和临床相关性
细菌学培养、PCR和免疫荧光法对牛尿液标本中的博格彼得森氏乳杆菌血清型Hardjo型Hardjo-bovis的检测敏感，但单一技术对每只受试动物不是最敏感的。因此，联合使用2种技术可获得最大的诊断灵敏度。
OBJECTIVE
To compare sensitivity and specificity of various polymerase chain reaction (PCR) assays for detection of Leptospira borgpetersenii serovar hardjo in bovine urine and to compare results of the optimal PCR assay with results of immunofluorescence, nucleic acid hybridization, and bacteriologic culture.
ANIMALS
6 heifers.
PROCEDURE
Heifers were exposed to serovar hardjo type hardjo-bovis by conjunctival instillation of 10(6) leptospires on 3 successive days. Urine samples were collected before and after infection. Sensitivity and specificity of 5 PCR assays were compared, to determine the optimal assay for use with bovine urine samples. The optimal PCR assay was then compared with results of bacteriologic culture, nucleic acid hybridization, and immunofluorescence.
RESULTS
A PCR assay with the best combination of specificity (100%) and sensitivity (91%) was selected for comparison with the other diagnostic tests. Sensitivity for nucleic acid hybridization was 55%, whereas sensitivity for bacteriologic culture and immunofluorescence was 89 to 93%.
CONCLUSIONS AND CLINICAL RELEVANCE
Bacteriologic culture, PCR, and immunofluorescence were sensitive for detection of L borgpetersenii serovar hardjo type hardjo-bovis in urine specimens of cattle, but a single technique was not the most sensitive for each animal tested. Therefore, the use of 2 techniques in combination is warranted for maximal sensitivity for diagnosis.