怎样提取猪瘟病毒的RNA

J Vet Diagn Invest 17:574–578 (2005)
Comparison of six RNA extraction methods for the detection of classical swine fever
virus by real-time and conventional reverse transcription–PCR
Ming Y. Deng,1 He Wang, Gordon B. Ward, Tammy R. Beckham, Thomas S. McKenna
Abstract. Six RNA extraction methods, i.e., RNAqueous kit, Micro-to-midi total RNA purification system,
NucleoSpin RNA II, GenElute mammalian total RNA kit, RNeasy mini kit, and TRIzol LS reagent, were
evaluated on blood and 7 tissues from pig infected with classical swine fever virus (CSFV). Each of the 6
extraction methods yielded sufficient RNA for positive results in a real-time reverse transcription–PCR (RTPCR)
for CSFV, and all RNA, except the one extracted from blood by TRIzol LS reagent, yielded positive
results in both a conventional RT-PCR for CSFV and a conventional RT-PCR for an endogenous gene encoding
b-actin. The RNA extracted from blood by TRIzol LS reagent became positive in both conventional RT-PCR
assays when it was diluted to 1:2, 1:4, or up to 1:64 in nuclease-free water. It is concluded that all 6 methods
are more or less useful for the detection of CSFV by real-time and conventional RT-PCR in swine blood and
tissues. However, some of the 6 reagents offer certain advantages not common to all 6 extraction procedures.
For example, RNA extracted by the TRIzol LS reagent constantly had the highest yield; that by the RNAqueous
kit had the highest A260/A280 ratio for almost all samples; and that by the NucleoSpin RNA II and the GenElute
mammalian total RNA kit was most likely to be free of contaminations with genomic DNA.

有专门的试剂盒可以提取,比如罗氏