Quantitative TaqMan® RT-PCR for the detection and differentiation of European and North American strains of procine reproductive and respiratory syndrome virus

被引:73
|
作者
Egli, C [1 ]
Thür, B [1 ]
Liu, LZ [1 ]
Hofmann, MA [1 ]
机构
[1] Inst Virol & Immunoprophylaxis, CH-3147 Mittelhausen, Switzerland
关键词
PRRSV; differentiation; RT-PCR; TaqMan (R);
D O I
10.1016/S0166-0934(01)00358-5
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Since two different types of porcine reproductive and respiratory syndrome virus (PRRSV), the European (EU) and the North American (US) strain, occur or coexist in European swine herds, their rapid and reliable detection and differentiation is essential for disease surveillance. A quantitative TaqMan (R) reverse transcription-polymerise chain reaction (RT-PCR) is described for PRRSV detection and strain differentiation. Sensitivity and specificity were compared with a conventional PRRSV RT-PCR and to the detection of both PRRSV types in cell cultures and both were found to be equal or superior to the reference methods. Reproducibility was tested and proved that the assay was very reliable. Standard dilutions included in each test allowed absolute quantitation of the amount of viral RNA. The TaqMan (R) assay described below is time-saving, easy to handle, exhibits a decreased risk of cross-contamination and is highly sensitive and specific. It is, therefore, considered to be a powerful tool for the rapid detection and differentiation of PRRSV. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:63 / 75
页数:13
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