Comparison of virus isolation and reverse transcription polymerase chain reaction assay for detection of bovine viral diarrhea virus in bulk milk tank samples
Comparison of virus isolation and reverse transcription polymerase chain reaction assay for detection of bovine viral diarrhea virus in bulk milk tank samples
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DOI：
10.1177/104063870001200219
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发表时间：
2000-03-01
影响因子：
1.5
通讯作者：
Dubovi, EJ
中科院分区：
文献类型：
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作者：
Renshaw, RW;Ray, R;Dubovi, EJ
使用逆转录酶聚合酶链反应（RT-PCR）试验筛选散装奶罐样品中的牛病毒性腹泻病毒（BVDV）已被证明是评价牛群中泌乳动物的敏感且经济的方法。该检测试剂盒能够检测一组数百头奶牛中是否存在单个持续感染动物。在3年时间内，采用RT-PCR试验结合经典病毒分离（VI）程序检测了来自97个农场的144份样本的BVDV，以测量技术的相对有效性。当用一群BVDV阴性动物的乳汁以1：600稀释单个持续感染动物的乳汁时，两种方法均可检出病毒。基于个体农场，BVDV感染的总体流行率为12.4%，2种检测之间的相关性为95.9%。在敏感性、特异性和周转时间方面，RT-PCR上级VI。然而，在17份VI阳性样本中，4份RT-PCR阴性。RT-PCR可能无法检出所有天然存在的BVDV分离株，因为它们在引物区域可能含有微小的序列变异。当单独使用时，VI和RT-PCR均适用于检测散装牛奶样品中的BVDV，但为了增加成功检测的可能性并提供针对试验污染的交叉检查，需要平行使用这两种方法。
The use of a reverse transcriptase polymerase chain reaction (RT-PCR) assay to screen bulk milk tank samples for bovine viral diarrhea virus (BVDV) has proven to be a sensitive and economical means to evaluate the lactating animals in a herd. The assay is capable of detecting the presence of a single persistently infected animal within a group of several hundred cows. Over a 3-year period, 144 samples from 97 farms were tested for BVDV using an RT-PCR assay in conjunction with a classical virus isolation (VI) procedure to measure the relative effectiveness of the techniques. Virus could be detected with both methods when the milk from a single persistently infected animal was diluted 1:600 with the milk from a herd of BVDV-negative animals. Based on individual farms, there was an overall prevalence of 12.4% BVDV infection, and the correlation between the 2 assays was 95.9%. In terms of sensitivity, specificity, and turnaround time, RT-PCR was superior to VI. However, of the 17 samples that were VI positive, 4 were RT-PCR negative. RT-PCR may not detect all naturally occurring BVDV isolates because they may contain minor sequence variations in the primer regions. VI and RT-PCR are both suitable for detection of BVDV in bulk milk samples when used independently, but to increase the probability of successful detection and to provide cross-checks against assay contamination, it is desirable to utilize both methods in parallel.